KR20220012840A - Co-agonists at GLP-1 and GIP receptors suitable for oral delivery - Google Patents

Abstract

Peptide co-agonists of human GLP-1 and GIP receptors suitable for oral delivery, including long-acting derivatives, and their medical use in the treatment and/or prophylaxis of obesity, diabetes and/or liver disease are described. do.

Description

Co-agonists at GLP-1 and GIP receptors suitable for oral delivery

The present invention relates to novel compounds that are agonists of the glucagon-like peptide 1 (GLP-1) receptor and the glucose-dependent insulinotropic polypeptide (GIP) receptor with an extended profile of action, suitable for oral administration to humans.

Incorporation by Reference in the Sequence Listing

This application is filed with a sequence listing in electronic format. The entire contents of the Sequence Listing are incorporated herein by reference.

Glucagon-like peptide 1 (GLP-1) is an enteroendocrine cell-derived hormone and is one of two prominent endogenous physiological incretins. GLP-1 improves glycemic control by stimulating glucose-dependent insulin secretion in response to nutrient (glucose), inhibits glucagon secretion from pancreatic alpha-cells, slows gastric emptying, and reduces food consumption, resulting in primary weight loss induce Glucose-dependent insulin-releasing polypeptide (GIP), another excellent incretin, improves glycemic control by stimulating insulin secretion in response to nutrients (fat, glucose). In addition, GIP appears to improve plasma lipid profiles and stimulate calcium accumulation in bones. In contrast to GLP-1, the incretin effect of GIP is severely reduced in patients with type 2 diabetes, but a recent study suggests that GIP efficiency may be restored after treatment in these patients and may improve glycemic control. do. Nevertheless, the role of GIP in regulating systemic metabolism beyond its direct effect in the endocrine pancreas is controversial, especially as it relates to the action of GIP in promoting an increase in body fat mass in animal models. These results promoted the belief that GIPR antagonism could improve body weight. Therefore, as a strategy for improving body weight, compounds that act at GIP receptors, in particular, whether agonists or antagonists are still the subject of intensive scientific investigation (Finan et al., TRENDS Mol Med, 2016, 22(5): 359-376; Killion et al., Endo Rev, 2020, 41(1): 1-21).

Prolonged GIP analogs have been shown to lower body weight and improve glycemic control, but are relatively less potent at lowering body weight than GLP-1 analogs in rodent models (Mroz et al., Mol Metab, 2019, 20: 51-62). In addition, GIP analogues induce weight loss by additive/homologous action with GLP-1 analogues in dual administration (Finan et al., Sci Transl Med, 2013, 5(209): 209ra151; Nørregaard et al., Diabes Obes Metab). , 2018, 20(1): 60-68), thus representing a suitable candidate for amplification of GLP-1-based pharmacology. GIPR action can be recruited as a non-overlapping partner for GLP-1R action, as a single molecule co-agonist amplifying the GLP-1 metabolic effect, the most prominent weight loss and glycemic control, as shown in preclinical animal models. (Finan et al., Sci Transl Med, 2013, 5(209): 209ra151; Coskun et al., Mol Metab, 2018, 18:3-14). Two different peptides with high potency dual incretin receptor action were conducted in a multi-dose clinical study. Clinical results demonstrated improvements in glycemic control and body weight beyond those achieved with doses similar to the benchmark GLP-1 specific agonist (Frias et al., Cell Metab, 2017, 26(2): 343-352; Frias et al. , Lancet, 2018, 392(10160): 2180-2193), which demonstrates a translational modality and therapeutic benefit of co-targeting GLP-1 and the GIP receptor.

Oral delivery of GLP-1 derivatives is a once-daily tablet of semaglutide and the penetration enhancer sodium N- [8-(2-hydroxyl) to improve the bioavailability and very low typical exposure of GLP-1 derivatives after oral administration. The form of benzoyl)amino]caprylate (SNAC) (Hedrington & Davis, Exp. Opin. Pharmacother. 2019, 20 (2): 133-141) has been investigated clinically.

GLP-1/GIP co-agents and their potential medical uses are described in WO 2010/011439, WO 2013/164483, WO 2014/192284, WO 2015/067715, WO 2015/022420, WO 2015/086728, WO 2015/086729, It is described in several patent applications such as WO 2016/111971, WO 2020/023386, US 9745360, US 2014/162945, and US 2014/0357552. Patent applications disclosing oral delivery of GLP-1 derivatives are described, for example, in WO 2011/080103, WO 2012/080471, WO 2013/189988, and WO 2019/149880.

However, to date, no co-agent product has been approved for marketing.

The present invention relates to single molecule co-agents comprising peptides and substituents that react with both human GLP-1 and GIP receptors with high potency and are suitable for daily oral administration to humans. This is achieved by combining specific peptide sequence variants and substituents via single site acylation with discrete fatty acids.

One aspect of the present invention relates to a peptide having the following amino acid sequence,

YX ₂ EGTX ₆ TSDYSX ₁₂ X ₁₃ LEX ₁₆ QAAX ₂₀ X ₂₁ FX ₂₃ X ₂₄ WLLX ₂₈ GGPX ₃₂ X ₃₃ X ₃₄ X ₃₅ X ₃₆ X ₃₇ X ₃₈ X ₃₉ (SEQ ID NO: 36);

It has an optional amide modification at the C-terminus,

From here;

X ₂ is Aib or A,

X ₆ is F or V,

X ₁₂ is I or Y,

X ₁₃ is Y, A, L or I;

X ₁₆ is K or E,

X ₂₀ is Q, R, E, H,

X ₂₁ is A or E,

X ₂₃ is I or V,

X ₂₄ is E, Q or N,

X ₂₈ is A or R,

X ₃₂ is E, S or absent;

X ₃₃ is S, K or absent;

X ₃₄ is G or absent;

X ₃₅ is A or absent,

X ₃₆ is P or absent;

X ₃₇ is P or absent;

X ₃₈ is P or absent;

X ₃₉ is S or absent.

In another aspect, the present invention relates to a peptide having the following amino acid sequence, or a pharmaceutically acceptable salt thereof,

YX ₂ EGTX ₆ TSDYSX ₁₂ X ₁₃ LEX ₁₆ QAAX ₂₀ X ₂₁ FX ₂₃ X ₂₄ WLLX ₂₈ GGPX ₃₂ X ₃₃ X ₃₄ X ₃₅ X ₃₆ X ₃₇ X ₃₈ X ₃₉ (SEQ ID NO: 36)

It has an optional amide modification at the C-terminus,

From here;

X ₂ is Aib or A,

X ₆ is F or V,

X ₁₂ is I or Y,

X ₁₃ is Y, A, L or I;

X ₁₆ is K or E,

X ₂₀ is Q, R, E, H,

X ₂₁ is A or E,

X ₂₃ is I or V,

X ₂₄ is E, Q or N,

X ₂₈ is A or R,

X ₃₂ is E, S or absent;

X ₃₃ is S, K or absent;

X ₃₄ is G or absent;

X ₃₅ is A or absent,

X ₃₆ is P or absent;

X ₃₇ is P or absent;

X ₃₈ is P or absent;

X ₃₉ is S or absent;

The substituent is attached to the peptide via a lysine (K) at position 16 or 33.

Another aspect of the invention relates to a process for the preparation of the GLP-1/GIP co-agonists described herein.

In a further aspect, the present invention relates to a pharmaceutical composition comprising a GLP-1/GIP co-agonist compound as described herein.

Another aspect of the invention relates to the medical use of the GLP-1/GIP co-agonists described herein.

In one aspect, the invention relates to the use of a GLP-1/GIP co-agonist as described herein for the prophylaxis or treatment of diabetes, obesity and/or liver disease.

In the following, Greek letters may be denoted by symbols or their corresponding names (eg, α = alpha; β = beta; ε = epsilon; γ = gamma; ω = omega, etc.). Also, the Greek letter μ can be denoted as “u” (eg μl = ul, or μM = uM).

GLP-1/GIP receptor co-agonists

The present invention relates to compounds that are GLP-1 receptors and GLP receptor agonists, also referred to as GLP-1/GIP receptor co-agonists or simply co-agonists.

The term “compound” is used herein to refer to a molecular entity, and thus a “compound” may have structural components that differ in addition to the minimum components defined for each compound in a group of compounds. This means that a compound may be a peptide or a derivative thereof, so long as it contains defined structural and/or functional components.

The term “compound” also relates to a pharmaceutically related form thereof, ie a compound as defined herein or a pharmaceutically acceptable salt, amide or ester thereof.

The term “analog” generally refers to a peptide having a sequence in which one or more amino acids are altered as compared to a reference amino acid sequence. An “analog” includes amino acid elongation at N-terminal and/or C-terminal positions and/or may include cleavage at N-terminal and/or C-terminal positions.

In general, amino acid residues can be identified by their full name, their one-letter code and/or their three-letter code. These three methods are completely equivalent.

Amino acids are molecules that contain an amino group and a carboxylic acid group, optionally containing one or more additional groups, often referred to as side chains.

The term "amino acid" includes proteinogenic (or natural) amino acids (among its 20 standard amino acids) as well as non-proteinogenic (or non-natural) amino acids. Proteinogenic amino acids are those that are naturally incorporated into proteins.

Standard amino acids are those encoded by the genetic code. Non-proteinogenic amino acids are not found in proteins or produced by standard cellular machinery (ie, may be subject to post-translational modifications). Non-limiting examples of non-proteinogenic amino acids include Aib (α-aminoisobutyric acid or 2-aminosisobutyric acid), norleucine, norvaline, as well as the D-isomer of proteinogenic amino acids.

Next, each amino acid of a peptide in which no optical isomer is recited will be understood to mean the L-isomer (unless otherwise specified).

The GLP-1/GIP receptor co-agonists described herein comprise or consist of a peptide and a substituent as defined below. In some embodiments, the peptide is a synthetic peptide generated to optimize activity at GLP-1 and GIP receptors. Compounds with appropriate receptor binding activity for both the GLP-1 receptor and the GIP receptor were identified as demonstrated in the Examples herein.

The compound further exhibits an extended half-life obtained by a substituent comprising a fatty acid group.

In some embodiments, the carboxy terminus of the peptide bears a -COOH group. In some embodiments, compounds may optionally include an amide group (C(=O)-NH ₂ ) at the C-terminus, which converts —OH to —NH ₂ , as seen in native Exendin-4. It is a naturally-occurring transformation that substitutes.

peptide

The GLP-1/GIP receptor co-agonists described herein comprise a peptide and a substituent as defined below, wherein the substituent is attached to the peptide backbone through an amino acid residue.

In some embodiments, the amino acid sequence of the peptide is

YX ₂ EGTX ₆ TSDYSX ₁₂ X ₁₃ LEX ₁₆ QAAX ₂₀ X ₂₁ FX ₂₃ X ₂₄ WLLX ₂₈ GX ₃₀ X ₃₁ X ₃₂ X ₃₃ X ₃₄ X ₃₅ X ₃₆ X ₃₇ X ₃₈ X ₃₉ (SEQ ID NO: 47)

It has an optional amide modification at the C-terminus,

From here;

X ₂ is Aib or A,

X ₆ is F or V,

X ₁₂ is I or Y,

X ₁₃ is Y, A, L or I;

X ₁₆ is K or E,

X ₂₀ is Q, R, E, H,

X ₂₁ is A or E,

X ₂₃ is I or V,

X ₂₄ is E, Q or N,

X ₂₈ is A or R,

X ₃₀ is G or absent;

X ₃₁ is P or absent,

X ₃₂ is E, S or absent;

X ₃₃ is S, K or absent;

X ₃₄ is G or absent;

X ₃₅ is A or absent,

X ₃₆ is P or absent;

X ₃₇ is P or absent;

X ₃₈ is P or absent;

X ₃₉ is S or absent.

In some embodiments, the amino acid sequence of the peptide is

YX ₂ EGTX ₆ TSDYSX ₁₂ X ₁₃ LEX ₁₆ QAAX ₂₀ X ₂₁ FX ₂₃ X ₂₄ WLLX ₂₈ GGPX ₃₂ X ₃₃ X ₃₄ X ₃₅ X ₃₆ X ₃₇ X ₃₈ X ₃₉ (SEQ ID NO: 36)

It has an optional amide modification at the C-terminus,

From here;

X ₂ is Aib or A,

X ₆ is F or V,

X ₁₂ is I or Y,

X ₁₃ is Y, A, L or I;

X ₁₆ is K or E,

X ₂₀ is Q, R, E, H,

X ₂₁ is A or E,

X ₂₃ is I or V,

X ₂₄ is E, Q or N,

X ₂₈ is A or R,

X ₃₂ is E, S or absent;

X ₃₃ is S, K or absent;

X ₃₄ is G or absent;

X ₃₅ is A or absent,

X ₃₆ is P or absent;

X ₃₇ is P or absent;

X ₃₈ is P or absent;

X ₃₉ is S or absent.

In one embodiment, X ₃₉ is absent. In one embodiment, X ₃₈ and X ₃₉ are absent. In one embodiment, X _37, X ₃₈ and X ₃₉ are absent. In one embodiment, X _36, X _37, X ₃₈ and X ₃₉ are absent. In one embodiment, X ₃₅ , X _{36 ,} X _{37 ,} X ₃₈ and X ₃₉ are absent. In one embodiment, X ₃₄ , X ₃₅ , X _{36 ,} X _{37 ,} X ₃₈ and X ₃₉ are absent. In one embodiment, X ₃₃ , X ₃₄ , X ₃₅ , X _{36 ,} X _{37 ,} X ₃₈ and X ₃₉ are absent. In one embodiment, X ₃₂ , X ₃₃ , X ₃₄ , X ₃₅ , X _{36 ,} X _{37 ,} X ₃₈ and X ₃₉ are absent. In one embodiment, X ₃₁ , X ₃₂ , X ₃₃ , X ₃₄ , X ₃₅ , X _{36 ,} X _{37 ,} X ₃₈ and X ₃₉ are absent. In one embodiment, X ₃₀ , X ₃₁ , X ₃₂ , X ₃₃ , X ₃₄ , X ₃₅ , X _{36 ,} X _{37 ,} X ₃₈ and X ₃₉ are absent. In further such embodiments, X ₃₂ X ₃₃ X ₃₄ X ₃₅ is SSGA. In further such embodiments, X ₃₂ X ₃₃ X ₃₄ X ₃₅ is ESGA. In further such embodiments, X ₃₂ X ₃₃ X ₃₄ X ₃₅ is SKGA. In a further embodiment thereof, the peptide has an amide modification at the C-terminus.

In one embodiment, the amino acid sequence of the peptide is

YX ₂ EGTX ₆ TSDYSX ₁₂ X ₁₃ LEX ₁₆ QAAX ₂₀ X ₂₁ FX ₂₃ X ₂₄ WLLX ₂₈ GGPSSGAPPPS (SEQ ID NO: 37)

From here;

X ₂ is Aib or A,

X ₆ is F or V,

X ₁₂ is I or Y,

X ₁₃ is Y, A, L or I;

X ₁₆ is K or E,

X ₂₀ is Q, R, E, H,

X ₂₁ is A or E,

X ₂₃ is I or V,

X ₂₄ is E, Q or N,

X ₂₈ is A or R.

In one embodiment, the amino acid sequence of the peptide is

YX ₂ EGTX ₆ TSDYSX ₁₂ X ₁₃ LEX ₁₆ QAAX ₂₀ X ₂₁ FX ₂₃ X ₂₄ WLLX ₂₈ GGPESGAPPPS (SEQ ID NO: 38)

From here;

X ₂ is Aib or A,

X ₆ is F or V,

X ₁₂ is I or Y,

X ₁₃ is Y, A, L or I;

X ₁₆ is K or E,

X ₂₀ is Q, R, E, H,

X ₂₁ is A or E,

X ₂₃ is I or V,

X ₂₄ is E, Q or N,

X ₂₈ is A or R.

In one embodiment, the amino acid sequence of the peptide is

YX ₂ EGTX ₆ TSDYSX ₁₂ X ₁₃ LEX ₁₆ QAAX ₂₀ X ₂₁ FX ₂₃ X ₂₄ WLLX ₂₈ GGPSKGAPPPS (SEQ ID NO: 39)

From here;

X ₂ is Aib or A,

X ₆ is F or V,

X ₁₂ is I or Y,

X ₁₃ is Y, A, L or I;

X ₁₆ is K or E,

X ₂₀ is Q, R, E, H,

X ₂₁ is A or E,

X ₂₃ is I or V,

X ₂₄ is E, Q or N,

X ₂₈ is A or R.

In one embodiment, X ₂ is Aib. In one embodiment, X ₂ is A. In one embodiment, X ₆ is F. In one embodiment, X ₆ is V. In one embodiment, X ₁₂ is I. In one embodiment, X ₁₂ is Y. In one embodiment, X ₁₃ is Y. In one embodiment, X ₁₃ is A. In one embodiment, X ₁₃ is L. In one embodiment, X ₁₃ is I. In one embodiment, X ₁₆ is K. In one embodiment, X ₁₆ is E. In one embodiment, X ₂₀ is Q. In one embodiment, X ₂₀ is R. In one embodiment, X ₂₀ is E. In one embodiment, X ₂₀ is H. In one embodiment, X ₂₁ is A. In one embodiment, X ₂₁ is E. In one embodiment, X ₂₃ is I. In one embodiment, X ₂₃ is V. In one embodiment, X ₂₄ is E. In one embodiment, X ₂₄ is Q. In one embodiment, X ₂₄ is N. In one embodiment, X ₂₈ is A. In one embodiment, X ₂₈ is R. In one embodiment, X ₃₀ is G. In one embodiment, X ₃₁ is P.

In one embodiment, X ₁₃ LEX ₁₆ QAAX ₂₀ X ₂₁ FX ₂₃ X ₂₄ is selected from the group consisting of: a peptide selected from the group consisting of LLEKQAAREFIN, LLEKQAAREFIE, LLEKQAAQEFIE and LLEEQAAREFIE. In one embodiment, X ₁₃ LEX ₁₆ QAAX ₂₀ X ₂₁ FX ₂₃ X ₂₄ is LLEKQAAREFIN. In one embodiment, X ₁₃ LEX ₁₆ QAAX ₂₀ X ₂₁ FX ₂₃ X ₂₄ is LLEKQAAREFIE. In one embodiment, X ₁₃ LEX ₁₆ QAAX ₂₀ X ₂₁ FX ₂₃ X ₂₄ is LLEKQAAQEFIE. In one embodiment, X ₁₃ LEX ₁₆ QAAX ₂₀ X ₂₁ FX ₂₃ X ₂₄ is LLEEQAAREFIE.

In further embodiments, the amino acid sequence of the peptide is SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 , 20, 21, 22, 23, 24, 25, 26, 27, 33, 34 and 35.

In one embodiment, the amino acid sequence of the peptide is any one of SEQ ID NOs: 10, 22 or 25.

In one embodiment, the amino acid sequence of the peptide is SEQ ID NO:25.

In one embodiment, the amino acid sequence of the peptide is SEQ ID NOs: 18, 20, 23, 24.

In one embodiment, the amino acid sequence of the peptide is SEQ ID NO: 33, 34 or 35.

In a further such embodiment, the peptide has an amide modification at the C-terminus.

derivative

In some embodiments, the GLP-1 and GIP receptor agonists comprise or consist of a substituent as defined below covalently bound to the peptide.

Such compounds may be referred to as derivatives of peptides, since they are obtained by covalently attaching a substituent to the peptide backbone.

One aspect of the present invention relates to a compound comprising a peptide and a substituent or a pharmaceutically acceptable salt thereof, and the amino acid sequence of the peptide is as follows,

YX ₂ EGTX ₆ TSDYSX ₁₂ X ₁₃ LEX ₁₆ QAAX ₂₀ X ₂₁ FX ₂₃ X ₂₄ WLLX ₂₈ GX ₃₀ X ₃₁ X ₃₂ X ₃₃ X ₃₄ X ₃₅ X ₃₆ X ₃₇ X ₃₈ X ₃₉ (SEQ ID NO: 47)

It has an optional amide modification at the C-terminus,

From here;

X ₂ is Aib or A,

X ₆ is F or V,

X ₁₂ is I or Y,

X ₁₃ is Y, A, L or I;

X ₁₆ is K or E,

X ₂₀ is Q, R, E, H,

X ₂₁ is A or E,

X ₂₃ is I or V,

X ₂₄ is E, Q or N,

X ₂₈ is A or R,

X ₃₀ is G or absent;

X ₃₁ is P or absent,

X ₃₂ is E, S or absent;

X ₃₃ is S, K or absent;

X ₃₄ is G or absent;

X ₃₅ is A or absent,

X ₃₆ is P or absent;

X ₃₇ is P or absent;

X ₃₈ is P or absent;

X ₃₉ is S or absent;

The substituent is attached to the peptide via a lysine (K) at position 16 or 33.

One embodiment of the present invention relates to a compound comprising a peptide and a substituent or a pharmaceutically acceptable salt thereof, and the amino acid sequence of the peptide is as follows,

YX ₂ EGTX ₆ TSDYSX ₁₂ X ₁₃ LEX ₁₆ QAAX ₂₀ X ₂₁ FX ₂₃ X ₂₄ WLLX ₂₈ GGPX ₃₂ X ₃₃ X ₃₄ X ₃₅ X ₃₆ X ₃₇ X ₃₈ X ₃₉ (SEQ ID NO: 36)

It has an optional amide modification at the C-terminus,

From here;

X ₂ is Aib or A,

X ₆ is F or V,

X ₁₂ is I or Y,

X ₁₃ is Y, A, L or I;

X ₁₆ is K or E,

X ₂₀ is Q, R, E, H,

X ₂₁ is A or E,

X ₂₃ is I or V,

X ₂₄ is E, Q or N,

X ₂₈ is A or R,

X ₃₂ is E, S or absent;

X ₃₃ is S, K or absent;

X ₃₄ is G or absent;

X ₃₅ is A or absent,

X ₃₆ is P or absent;

X ₃₇ is P or absent;

X ₃₈ is P or absent;

X ₃₉ is S or absent;

The substituent is attached to the peptide via a lysine (K) at position 16 or 33.

In a further embodiment, a peptide may be defined as described above.

substituent

In one embodiment, a substituent described herein is attached to a peptide described herein via a lysine (K) residue at position 16 or 33.

In one embodiment, when the aforementioned lysine is included in position 16 or 33, the substituent is attached to the peptide via the epsilon-amino group of lysine (K).

In one embodiment, the substituent is a chemical structure covalently attached to a peptide capable of forming a non-covalent complex with plasma albumin, thereby facilitating circulation of the co-agent with the bloodstream, and also causing the complex of co-agent and albumin to elongate. It has the effect of prolonging the time of action of the co-agent due to the fact that it is removed only slowly by cleaning.

In some embodiments, the substituent comprises a fatty acid group. In this embodiment, the fatty acid group comprises a carbon chain containing at least 8 consecutive —CH ₂ —groups. In one embodiment, the fatty acid group has at least 10 consecutive —CH ₂ —groups, such as at least 12 consecutive —CH ₂ —groups, at least 14 consecutive —CH ₂ —groups, at least 16 consecutive —CH ₂ —groups, or at least contains 18 consecutive —CH ₂ —groups.

In one embodiment, the fatty acid group comprises 8 to 20 consecutive —CH ₂ —groups. In one embodiment, the fatty acid group comprises 10 to 18 consecutive —CH ₂ —groups. In one embodiment, the fatty acid group comprises 12 to 18 consecutive —CH ₂ —groups. In one embodiment, the fatty acid group comprises 14 to 18 consecutive —CH ₂ —groups.

In some embodiments, a substituent consists of several elements, such as a protractor element and one or more linker elements. In one embodiment, the term “extending group” is used to describe a fatty acid group responsible for extending the half-life of a compound, as the terminus of a substituent.

In one embodiment, the extender (Prot) is

Formula 1: HOOC-(CH ₂ ) _n -CO- ^* , wherein n is an integer ranging from 8 to 20, which is referred to as a C(n+2) diacid, or

(여기에서, n은 8 내지 20 범위의 정수임)로서 정의될 수 있다.Formula 1b:

(wherein n is an integer ranging from 8 to 20).

In one embodiment, the substituent further comprises one or more linker elements. In some embodiments, the linker elements are linked to each other and to the extender by amide bonds and may be referred to as “Z” (see below for details).

As further defined herein, the number of linker elements is up to three, which may be referred to as -Z1-Z2-Z3-, wherein Z1 is linked to an extender (Prot) and the last Z element is linked to a peptide linked, in which case the substituent may be referred to as Prot-Z1-Z2-Z3-. Thus, the symbol "*" above indicates the point of attachment to Z1, which is nitrogen when bonded via an amide bond. In one embodiment, when Z 1 is a bond (see below), the symbol * indicates the point of attachment to the nitrogen of the neighboring Z element.

In one embodiment, the substituent is defined as Prot-Z1-Z2-Z3, wherein Prot- is selected from Formula 1, Formula 1b, and n is an integer ranging from 16 to 20.

In certain embodiments, n in Formula 1 or Formula 1b is 14, 15, 16, 17, 18, 19, or 20.

In certain embodiments, n in Formula 1 or Formula 1b is 14, 15, 16, 17 or 18.

In certain embodiments, n in Formula 1 or Formula 1b is 14, 16, or 18.

In certain embodiments, n in Formula 1 or Formula 1b is 16, 17, 18, 19 or 20.

In certain embodiments, n in Formula 1 or Formula 1b is 16, 18 or 20.

In certain embodiments, n in Formula 1 or Formula 1b is 18 or 20.

In certain embodiments, the Protractor (Prot) is a C16 discrete or a C18 discrete.

In certain embodiments, the extender (Prot) is a C18 diacid or a C20 diacid.

In certain embodiments, the extender (Prot) is a C16, C18 diacid or C20 diacid.

As used herein, the term “bond” refers to a covalent bond. When the linker element of Z1 to Z3 is defined as a bond, this is equivalent to the absence of the linker element described above. The following indication that any one of Z1 to Z3 is a bond can also be read as the absence of any one of Z1 to Z3, and is covalently bonded to the next Z element where the previous Z element is not "bonded" (or absent) do.

In some embodiments, linker elements Z1-Z3 are Glu, γGlu (also referred to as gamma Glu or gGlu, defined as ^* -NH-CH-(COOH)-CH ₂ -CH ₂ -CO- ^* ), ε-Lys amino acid-like moieties such as (also referred to as epsilon Lys or eLys, defined as ^* -NH-(CH ₂ ) ₄ -CH(NH ₂ )-CO- ^* ), Ser, Ala, Thr, Ado, Aeep and Aeeep and chemical moieties capable of forming amide bonds, including additional moieties described below.

In one embodiment, Z1 is selected from γGlu, Glu, or a bond.

In one embodiment, Z1 is γGlu.

In one embodiment, Z2 and Z3 are independently selected from each other from Glu, ε-Lys, γGlu, Gly, Ser, Ala, Thr, Ado, Aeep, Aeeep and a bond.

Glu, Gly, Ser, Ala, and Thr are amino acid residues well known in the art.

ε-Lys is defined by the formula 2: ^* -NH-(CH ₂ ) ₄ -CH(NH ₂ )-CO- ^* , which is also

로 나타낼 수 있다.Formula 2b:

can be expressed as

γGlu is defined by the formula 3: ^* -NH-CH(COOH)-(CH ₂ ) ₂ -CO- ^* , which is also

로 나타낼 수 있다.Formula 3b:

can be expressed as

Ado is defined by the formula 4: ^* -NH-(CH ₂ ) ₂ -O-(CH ₂ ) ₂ -O-CH ₂ -CO- ^* , which is referred to as 8-amino-3,6-dioxaoctanoic acid can be, which is also

로 나타낼 수 있다.Formula 4b:

can be expressed as

Aeep is defined by the formula 5: ^* NH-CH ₂ CH ₂ OCH ₂ CH ₂ OCH ₂ CH ₂ CO ^* , which is also

로 나타낼 수 있다.Formula 5b:

can be expressed as

Aeeep is defined by the formula 6: ^* NH-CH ₂ CH ₂ OCH ₂ CH ₂ OCH ₂ CH ₂ OCH ₂ CH ₂ CO ^* , which is also

로 나타낼 수 있다.Formula 6b:

can be expressed as

In one embodiment, Z2 and Z3 are independently selected from one another from Glu, ε-Lys, γGlu, Gly, Ala, Ado, Aeep, Aeeep and a bond.

In one embodiment, Z2 and Z3 are independently selected from each other from Glu, ε-Lys, γGlu, Gly, Ala, Ado and a bond.

In one embodiment, Z2 and Z3 are independently selected from each other from Glu, ε-Lys, γGlu, Gly, Ado and a bond.

In one embodiment, Z2 and Z3 are independently selected from each other from ε-Lys, γGlu, Gly, Ado and a bond.

In one embodiment, Z2 and Z3 are independently selected from each other from ε-Lys, γGlu, Ado and a bond.

In one embodiment, Z2 and Z3 are ε-Lys or Ado.

In one embodiment, Z2 and Z3 are Ado.

In one embodiment, Z2 and Z3 are ε-Lys.

In one embodiment, the substituents are selected from substituents A, B, C, D and E as defined below.

In one embodiment, the substituents are selected from substituents A, B and C defined as follows:

In some embodiments, the substituent is covalently attached to the lysine residue of the co-agent by acylation, i.e., via an amide bond formed between the carboxylic acid group of the substituent and the epsilon-amino group of the lysine residue.

In one embodiment, the substituent is covalently attached to the lysine residue at position 16 of the peptide backbone by acylation, ie via an amide bond formed between the carboxylic acid group of the substituent and the epsilon-amino group of the lysine residue.

In one embodiment, the substituent is covalently attached to the lysine residue at position 33 of the peptide backbone by acylation, ie via an amide bond formed between the carboxylic acid group of the substituent and the epsilon-amino group of the lysine residue.

Co-agents may exist in different stereoisomeric forms having the same molecular formula and binding atom sequence, but only the three-dimensional orientation of their atoms in space may differ. The stereoisomerism of the exemplified co-agents is indicated in the Experimental section by name and structure, using standard nomenclature. Unless otherwise stated, the present invention relates to all stereoisomeric forms of the embodied derivatives.

functional receptor activation activity

The functional activity of a GLP-1/GIP receptor agonist as described herein can be tested in vitro as described herein in Example 2.

The half effective concentration (EC ₅₀ ) generally refers to the concentration that induces a response at half the baseline and half the maximum with reference to the dose response curve. EC ₅₀ is used as a measure of the potency of a compound and represents the concentration at which 50% of the compound's maximal effect is observed.

Thus, the in vitro potency of a compound can be determined as described herein and the EC ₅₀ can be determined. The lower the EC ₅₀ value, the better the efficacy.

To characterize these compounds, it may be more appropriate to consider the efficacy in vitro compared to the natural hormone of each receptor.

In vitro efficacy can be determined, for example, in a medium containing a membrane expressing the appropriate GLP-1 and/or GIP receptor and/or in an assay using whole cells expressing the appropriate GLP-1 and/or GIP receptor. .

For example, a functional response of a human GLP-1 and/or GIP receptor is measured in a reporter gene assay, eg, in a stably transfected BHK cell line, wherein the cell line expresses a human GLP-1 and/or GIP receptor. and DNA for a cAMP response element (CRE) bound to a promoter and a gene for firefly luciferase (CRE luciferase). When cAMP is produced as a result of activation of GLP-1 and/or GIP receptors, this in turn leads to expression of luciferase. Luciferase can be determined by adding luciferin, which is enzymatically converted to oxyluciferin and produces bioluminescence, which is measured as a reporter of efficacy in vitro. An example of such an assay is described in Example 2 described herein. It is also important to recognize that receptor activity may be affected by the presence or absence of human serum albumin (HSA) in the assay medium, as compounds may contain substituents designed to bind to albumin. A decrease in potency of a compound in the presence of HSA, expressed as an increase in EC ₅₀ compared to EC ₅₀ in the absence of HSA, indicates an interaction of the compound with HSA and is predicted to delay the time of action in vivo.

In one embodiment, the compound has a potent in vitro effect on activating human GLP-1 and GIP receptors.

In one embodiment, the compound, when performed without HSA, is capable of activating human GLP-1 and GIP receptors in vitro with an EC ₅₀ of less than 20 pM in a CRE luciferase reporter assay as described in Example 2 herein. have.

In one embodiment, the compound is at a human GLP-1 and GIP receptor determined using the method of Example 2 corresponding to an EC ₅₀ of 100 pM or less, more preferably less than 50 pM, or most preferably less than 20 pM. It has in vitro potency.

In one embodiment, the EC ₅₀ in the human GLP-1 and GIP receptor assay is 1 to 25 pM, such as 1 to 20 pM, such as 1 to 15 pM, or such as 1 to 10 pM.

Pharmacokinetic properties

The pharmacokinetic properties of co-acting compounds can be further determined in vivo through pharmacokinetic (PK) studies. Animal models such as mouse, rat, monkey, dog, or pig can be used to perform this characterization.

In these studies, animals are usually administered intravenously, subcutaneously (sc) or orally (po) in a dosage form that involves a single dose of the drug. Blood samples are taken at predefined time points after dosing, and samples are analyzed for drug concentration using an associated quantitative assay. Based on these measurements, the time-plasma concentration profile for the study compound is plotted and a so-called non-compartmental pharmacokinetic analysis of the data is performed. An important parameter is the terminal half-life, as a long half-life indicates that less frequent dosing of the compound may be possible. The in vivo terminal half-life (t ^1/2 ) can be determined using appropriate models, eg after iv administration in minipigs as described in Example 3; or after po administration in the dogs described in Example 4.

In one embodiment, the terminal half-life is the in vivo half-life (t ^1/2 ) in minipigs following iv administration, eg, as described in Example 3 herein.

In one embodiment, the terminal half-life in minipigs is at least 24 hours, such as at least 30 hours, or such as at least 40 hours.

In one embodiment, the terminal half-life is the in vivo half-life (t ^1/2 ) in the dog after po administration, eg, as described in Example 4 herein.

In one embodiment, the terminal half-life in the dog is at least 24 hours, such as at least 40 hours, or such as at least 50 hours.

pharmaceutically acceptable salts

In some embodiments, the co-agent as described herein is in the form of a pharmaceutically acceptable salt. Salts are formed, for example, by a chemical reaction between a base and an acid (eg, 2NH ₃ + H ₂ SO ₄ → (NH ₄ ) ₂ SO ₄ ). A salt may be a basic salt, an acidic salt, or neither (ie, a neutral salt). Basic salts produce hydroxide ions and acid salt hydronium ions in water. Salts of co-agents may be formed with anionic groups or addition cations or anions between cationic groups, respectively. These groups may be located within the substituents of peptides and/or derivatives. Non-limiting examples of anionic groups include any free carboxylic acid group in a substituent, if any, in a peptide. The peptide may comprise a free carboxylic acid group at the C-terminus and, if present, any free carboxylic acid group of internal acidic amino acid residues such as Asp and Glu.

Non-limiting examples of cationic groups include any free amino group in a substituent, if any, in a peptide. The peptide may comprise a free amino group at the N-terminus and, if present, any free amino group of internal basic amino acid residues such as His, Arg and Lys.

In certain embodiments, the peptide or derivative is in the form of a pharmaceutically acceptable salt.

production process

Co-agents can be generated, for example, by classical peptide synthesis, such as by solid-phase peptide synthesis using t-Boc or Fmoc chemistry or other well-established techniques (eg, Greene and Wuts). , "Protective Groups in Organic Synthesis", John Wiley & Sons, 1999; Florencio Zaragoza Doerwald, "Organic Synthesis on solid Phase", Wiley-VCH Verlag GmbH, 2000, and edited by WC Chan and PD White, "Fmoc Solid Phase Peptide Synthesis" ", Oxford University Press, 2000).

Alternatively, the compound can be produced by recombinant methods, i.e., by culturing a host cell capable of expressing the peptide in an appropriate nutrient medium containing a DNA sequence encoding the peptide sequence and under conditions allowing expression of the peptide. have. Non-limiting examples of suitable host cells for expression of these peptides are mammalian BHK or CHO cell lines, including E. coli cell lines, Saccharomyces cerevisiae cell lines.

Co-agents comprising non-natural amino acids and/or covalently attached substituents can be produced as described in the experimental part.

Specific examples of methods for preparing multiple co-agents are included in the experimental part.

Another aspect of the invention relates to a method for making the peptides described herein.

Another aspect of the invention relates to a process for the preparation of the GLP-1/GIP co-agonists described herein.

In one embodiment, a method for preparing a compound as described herein comprises the step of solid phase peptide synthesis. Substituents can be constructed sequentially as part of solid phase peptide synthesis, or separately produced and attached via lysine residues after peptide synthesis.

In one embodiment, the compound is produced by a two-step process whereby two peptide fragments are linked after attachment of a substituent to one of the peptide fragments.

pharmaceutical composition

In a further aspect, the present invention relates to a pharmaceutical composition comprising a GLP-1/GIP receptor co-agonist as described herein. A composition comprising a compound of the present disclosure or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient may be prepared as known in the art.

The term “excipient” broadly refers to any ingredient other than the active therapeutic ingredient(s). An excipient may be an inert substance, an inert substance, and/or a pharmaceutically inert substance. Excipients can be used for various purposes, for example, as carriers, carriers, fillers, binders, lubricants, glidants, disintegrants, flow control agents, crystallization inhibitors, solubilizers, stabilizers, colorants, flavoring agents, surfactants, emulsifiers or these agents. can be used as a combination of and/or can be used to improve absorption of active substances. The amount of each excipient used may vary within the ordinary skill of the art. Techniques and excipients that may be used to formulate oral dosage forms are described in the Handbook of Pharmaceutical Excipients (e.g., 8th ed., edited by Sheskey et al., American Pharmaceuticals Association and Pharmaceutical Press, publications department of the Royal Pharmaceutical Society of Great Britain). (2017), and one of later editions); and Remington: The Science and Practice of Pharmacy (eg, 22nd ed., edited by Remington and Allen, Pharmaceutical Press (2013), and one of the later editions).

In one embodiment, the pharmaceutical composition may be in a solid dosage form, for example, a freeze-dried or spray-dried composition, which may be used as such, or a solvent and/or diluent is added thereto by a physician or patient prior to use. In a further embodiment, the pharmaceutical composition comprises the active ingredient, a salt of N- [8-(2 - hydroxybenzoyl ) amino]caprylate, and known in the art, for example WO 2012/080471, WO 2013/189988, or a solid dosage form consisting of one or more additional excipients using one or more of the dosage forms described in WO 2019/149880.

Alternatively, the pharmaceutical composition is a liquid formulation, such as an aqueous formulation. Liquid compositions suitable for injection can be prepared using conventional techniques in the pharmaceutical industry, which involve properly dissolving and mixing the ingredients to obtain the desired final product. Thus, according to one procedure, a GLP-1/GIP co-agonist as described herein is dissolved in an appropriate buffer at an appropriate pH. The composition is sterilized, for example, by aseptic filtration.

pharmaceutical indications

Another aspect of the invention relates to the use of a GLP-1/GIP receptor co-agonist compound as described herein as a medicament.

In one embodiment, the compounds described herein are for use in the medical treatment of:

(i) prevention of all forms of diabetes, such as hyperglycemia, type 2 diabetes, impaired glucose tolerance, type 1 diabetes, non-insulin dependent diabetes mellitus, MODY (adult onset diabetes in juveniles), gestational diabetes and/or reduction of HbA1C; / or treatment;

(ii) delaying or preventing the progression of a diabetic disease, such as the progression of type 2 diabetes, delaying the progression of impaired glucose tolerance (IGT) to insulin-demanding type 2 diabetes, delaying or preventing insulin resistance, and/or non-insulin delay of progression of Demanding Type 2 Diabetes to Insulin Requiring Type 2 Diabetes;

(iii) prevention and/or treatment of eating disorders (eg obesity), for example, by reducing food intake, weight loss, suppression of appetite, induction of satiety; treatment or prevention of binge eating disorder, bulimia nervosa and/or obesity induced by the administration of psychotropic drugs or steroids; decrease in gastric motility; delayed gastric emptying; increased physical mobility; and/or prevention and/or treatment of obesity comorbidities such as osteoarthritis and/or urinary incontinence;

(iv) weight maintenance after successful weight loss (either drug-induced or by diet control and exercise) - ie, prevention of weight gain after successful weight loss;

(v) the prevention and/or treatment of liver disorders such as hepatic steatosis, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), liver inflammation or fatty liver, and the like.

In one embodiment, the compound is for use in a method for the prevention and/or treatment of diabetes and/or obesity.

In one embodiment, the compound is for use in a method of treatment of diabetes and/or obesity.

In one embodiment, the compound is for use in a method of treating or preventing type 2 diabetes.

In one embodiment, the compound is for use in a method of treating type 2 diabetes.

In one embodiment, the compound is for use in a method of treating or preventing obesity.

In one embodiment, the compound is for use in a method of treating obesity.

In one embodiment, the compound is for use in a method of weight management. In one embodiment, the compound is for use in a method of reducing appetite. In one embodiment, the compound is for use in a method of reducing food intake.

implementation

1. A compound comprising a peptide and a substituent or a pharmaceutically acceptable salt thereof, wherein the amino acid sequence of the peptide is as follows,

YX ₂ EGTX ₆ TSDYSX ₁₂ X ₁₃ LEX ₁₆ QAAX ₂₀ X ₂₁ FX ₂₃ X ₂₄ WLLX ₂₈ GX ₃₀ X ₃₁ X ₃₂ X ₃₃ X ₃₄ X ₃₅ X ₃₆ X ₃₇ X ₃₈ X ₃₉ (SEQ ID NO: 47)

It has an optional amide modification at the C-terminus,

From here;

X ₂ is Aib or A,

X ₆ is F or V,

X ₁₂ is I or Y,

X ₁₃ is Y, A, L or I;

X ₁₆ is K or E,

X ₂₀ is Q, R, E, H,

X ₂₁ is A or E,

X ₂₃ is I or V,

X ₂₄ is E, Q or N,

X ₂₈ is A or R,

X ₃₀ is G or absent;

X ₃₁ is P or absent,

X ₃₂ is E, S or absent;

X ₃₃ is S, K or absent;

X ₃₄ is G or absent;

X ₃₅ is A or absent,

X ₃₆ is P or absent;

X ₃₇ is P or absent;

X ₃₈ is P or absent;

X ₃₉ is S or absent;

the substituent is attached to the peptide through a lysine (K) at position 16 or 33;

A compound or a pharmaceutically acceptable salt thereof.

2. The method of embodiment 1, comprising a peptide and a substituent, wherein the amino acid sequence of the peptide is as follows,

YX ₂ EGTX ₆ TSDYSX ₁₂ X ₁₃ LEX ₁₆ QAAX ₂₀ X ₂₁ FX ₂₃ X ₂₄ WLLX ₂₈ GGPX ₃₂ X ₃₃ X ₃₄ X ₃₅ X ₃₆ X ₃₇ X ₃₈ X ₃₉ (SEQ ID NO: 36)

It has an optional amide modification at the C-terminus,

From here;

X ₂ is Aib or A,

X ₆ is F or V,

X ₁₂ is I or Y,

X ₁₃ is Y, A, L or I;

X ₁₆ is K or E,

X ₂₀ is Q, R, E, H,

X ₂₁ is A or E,

X ₂₃ is I or V,

X ₂₄ is E, Q or N,

X ₂₈ is A or R,

X ₃₂ is E, S or absent;

X ₃₃ is S, K or absent;

X ₃₄ is G or absent;

X ₃₅ is A or absent,

X ₃₆ is P or absent;

X ₃₇ is P or absent;

X ₃₈ is P or absent;

X ₃₉ is S or absent;

the substituent is attached to the peptide through a lysine (K) at position 16 or 33;

A compound or a pharmaceutically acceptable salt thereof.

3. The compound of embodiment 1, wherein X _36, X _37, X ₃₈ and X ₃₉ are absent.

4. The compound of embodiment 1, wherein X ₃₄ , X ₃₅ , X ₃₆ , X ₃₇ , X ₃₈ and X ₃₉ are absent.

5. The compound of embodiment 1, wherein X ₃₂ , X ₃₃ , X ₃₄ , X ₃₅ , X _{36 ,} X _{37 ,} X ₃₈ and X ₃₉ are absent.

6. The compound of embodiment 1, wherein X ₃₀ , X ₃₁ , X ₃₂ , X ₃₃ , X ₃₄ , X ₃₅ , X _{36 ,} X _{37 ,} X ₃₈ and X ₃₉ are absent.

7. The compound according to any one of embodiments 1 to 6, wherein the peptide has an amide modification at the C-terminus.

8. The compound according to any one of embodiments 1, 2 or 7, wherein X ₃₂ X ₃₃ X ₃₄ X ₃₅ is selected from the group consisting of SSGA, ESGA and SKGA.

9. The compound of any one of embodiments 1, 2 or 7, wherein X ₃₂ X ₃₃ X ₃₄ X ₃₅ is SSGA.

10. The compound of any one of embodiments 1, 2 or 7, wherein X ₃₂ X ₃₃ X ₃₄ X ₃₅ is ESGA.

11. The compound according to any one of embodiments 1, 2 or 7, wherein X ₃₂ X ₃₃ X ₃₄ X ₃₅ is SKGA.

12. The amino acid sequence of embodiment 1, wherein the peptide is

YX ₂ EGTX ₆ TSDYSX ₁₂ X ₁₃ LEX ₁₆ QAAX ₂₀ X ₂₁ FX ₂₃ X ₂₄ WLLX ₂₈ GGPSSGAPPPS (SEQ ID NO: 37)

From here;

X ₂ is Aib or A,

X ₆ is F or V,

X ₁₂ is I or Y,

X ₁₃ is Y, A, L or I;

X ₁₆ is K or E,

X ₂₀ is Q, R, E, H,

X ₂₁ is A or E,

X ₂₃ is I or V,

X ₂₄ is E, Q or N,

X ₂₈ is A or R;

compound.

13. The amino acid sequence of embodiment 1, wherein the peptide is

YX ₂ EGTX ₆ TSDYSX ₁₂ X ₁₃ LEX ₁₆ QAAX ₂₀ X ₂₁ FX ₂₃ X ₂₄ WLLX ₂₈ GGPESGAPPPS (SEQ ID NO: 38)

From here;

X ₂ is Aib or A,

X ₆ is F or V,

X ₁₂ is I or Y,

X ₁₃ is Y, A, L or I;

X ₁₆ is K or E,

X ₂₀ is Q, R, E, H,

X ₂₁ is A or E,

X ₂₃ is I or V,

X ₂₄ is E, Q or N,

X ₂₈ is A or R;

compound.

14. The amino acid sequence of embodiment 1, wherein the peptide is

YX ₂ EGTX ₆ TSDYSX ₁₂ X ₁₃ LEX ₁₆ QAAX ₂₀ X ₂₁ FX ₂₃ X ₂₄ WLLX ₂₈ GGPSKGAPPPS (SEQ ID NO: 39)

From here;

X ₂ is Aib or A,

X ₆ is F or V,

X ₁₂ is I or Y,

X ₁₃ is Y, A, L or I;

X ₁₆ is K or E,

X ₂₀ is Q, R, E, H,

X ₂₁ is A or E,

X ₂₃ is I or V,

X ₂₄ is E, Q or N,

X ₂₈ is A or R;

compound.

15. The compound according to any one of embodiments 1 to 14, wherein X ₁₃ LEX ₁₆ QAAX ₂₀ X ₂₁ FX ₂₃ X ₂₄ is selected from the group consisting of LLEKQAAREFIN, LLEKQAAREFIE, LLEKQAAQEFIE and LLEEQAAREFIE.

16. The compound of embodiment 1, wherein the amino acid sequence of the peptide is any one of SEQ ID NOs: 1-27 or 33-35.

17. The compound of embodiment 16, wherein the peptide has an amide modification at the C-terminus.

18. The compound according to any one of embodiments 1 to 17, wherein the compound comprises human GLP-1 and GIP in vitro with an EC ₅₀ of less than 20 pM when measured without HSA in a CRE luciferase reporter assay as described in example 2 A compound that activates a receptor.

19. The compound according to any one of embodiments 1 to 18, which has a half-life in minipigs of at least 35 hours.

20. The compound of any one of embodiments 1-19, wherein the substituent is attached via 16Lys.

21. The compound of any one of embodiments 1-20, wherein the substituent is attached via 33Lys.

22. The compound of any one of embodiments 1-21, wherein the substituent comprises at least one extension group.

23. The compound of embodiment 22, wherein the extending group is a fatty acid group.

24. The extender of embodiment 23, wherein the extending group is a diacid defined by formula 1: HOOC-(CH ₂ ) _n —CO—, wherein n is an integer ranging from 8 to 20, eg, n=14, 16 or 18, compound.

25. The extender of embodiment 23, wherein the extending group is a diacid defined by formula 1: HOOC-(CH ₂ ) _n —CO—, wherein n is an integer ranging from 8 to 20, eg, n=16 or 18 , compound.

26. The compound according to any one of embodiments 1 to 25, wherein the substituent comprises at least one linker element.

27. The compound of embodiment 26, wherein the substituent comprises up to 3 linker elements.

28. The compound of embodiment 27, wherein the substituent comprises up to three linker elements, designated -Z1-Z2-Z3-, wherein -Z1- is linked to the extender and -Z3- is linked to the peptide.

29. according to any one of embodiments 1-28, the substituent is Prot-Z1-Z2-Z3-,

From here,

Prot is a C16-C20 diacid,

Z1 is γGlu or a bond,

Z2 is εLys, γGlu or Ado,

Z3 is εLys or Ado;

compound.

30. The compound of embodiment 29, wherein -Z1- is -γGlu-.

31. The compound of embodiment 29 or 30, wherein -Z2-Z3- is -Ado-Ado-.

32. The compound of embodiment 29, 30 or 31, wherein -Z2-Z3- is -εLys-εLys-.

33. The substituent of any of embodiments 1-17, wherein the substituents are:

A compound selected from the group consisting of.

34. The substituent of any of embodiments 1-17, wherein the substituents are:

A compound selected from the group consisting of.

35. The compound of any one of embodiments 1-17, wherein the substituent is attached to the peptide via the epsilon-amino group of lysine (K).

36. The compound of embodiment 1, wherein

A compound selected from the group consisting of.

37. The compound of clause 1, wherein

A compound selected from the group consisting of.

38. The compound of clause 1, wherein

A compound selected from the group consisting of.

39. The compound of embodiment 1, wherein

Phosphorus, compound.

40. The compound of embodiment 1, wherein

Phosphorus, compound.

41. A compound according to any one of embodiments 1 to 40, for use as a medicament.

42. A pharmaceutical composition comprising a compound according to any one of embodiments 1 to 40.

43. The composition of embodiment 42, wherein the composition is an aqueous liquid.

44. The composition of embodiment 42, wherein the composition is a solid composition.

45. The pharmaceutical composition according to any one of embodiments 42 to 44, for use in the treatment and/or prevention of diabetes and/or obesity.

46. of embodiments 42 to 44 for use in the prophylaxis and/or treatment of liver disorders such as hepatic steatosis, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), liver inflammation and/or fatty liver. The pharmaceutical composition according to any one of them.

47. A method for preventing and/or treating diabetes and/or obesity, comprising administering to a patient a pharmaceutically active amount of a compound according to any one of embodiments 1 to 40.

48. A method for the prophylaxis and/or treatment of liver disorders such as hepatic steatosis, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), liver inflammation and/or fatty liver, according to any one of embodiments 1 to 40. A method comprising administering a pharmaceutically active amount of a compound according to one.

49. A peptide, wherein the amino acid sequence of the peptide is:

YX ₂ EGTX ₆ TSDYSX ₁₂ X ₁₃ LEX ₁₆ QAAX ₂₀ X ₂₁ FX ₂₃ X ₂₄ WLLX ₂₈ GX ₃₀ X ₃₁ X ₃₂ X ₃₃ X ₃₄ X ₃₅ X ₃₆ X ₃₇ X ₃₈ X ₃₉ (SEQ ID NO: 47)

It has an optional amide modification at the C-terminus,

From here;

X ₂ is Aib or A,

X ₆ is F or V,

X ₁₂ is I or Y,

X ₁₃ is Y, A, L or I;

X ₁₆ is K or E,

X ₂₀ is Q, R, E, H,

X ₂₁ is A or E,

X ₂₃ is I or V,

X ₂₄ is E, Q or N,

X ₂₈ is A or R,

X ₃₀ is G or absent;

X ₃₁ is P or absent,

X ₃₂ is E, S or absent;

X ₃₃ is S, K or absent;

X ₃₄ is G or absent;

X ₃₅ is A or absent,

X ₃₆ is P or absent;

X ₃₇ is P or absent;

X ₃₈ is P or absent;

X ₃₉ is S or absent;

peptide.

50. A peptide, wherein the amino acid sequence of the peptide is:

YX ₂ EGTX ₆ TSDYSX ₁₂ X ₁₃ LEX ₁₆ QAAX ₂₀ X ₂₁ FX ₂₃ X ₂₄ WLLX ₂₈ GGPX ₃₂ X ₃₃ X ₃₄ X ₃₅ X ₃₆ X ₃₇ X ₃₈ X ₃₉ (SEQ ID NO: 36)

It has an optional amide modification at the C-terminus,

From here;

X ₂ is Aib or A,

X ₆ is F or V,

X ₁₂ is I or Y,

X ₁₃ is Y, A, L or I;

X ₁₆ is K or E,

X ₂₀ is Q, R, E, H,

X ₂₁ is A or E,

X ₂₃ is I or V,

X ₂₄ is E, Q or N,

X ₂₈ is A or R,

X ₃₂ is E, S or absent;

X ₃₃ is S, K or absent;

X ₃₄ is G or absent;

X ₃₅ is A or absent,

X ₃₆ is P or absent;

X ₃₇ is P or absent;

X ₃₈ is P or absent;

X ₃₉ is S or absent;

peptide.

51. The peptide of embodiment 49, wherein X _36, X _37, X ₃₈ and X ₃₉ are absent.

52. The peptide of embodiment 49, wherein X ₃₄ , X ₃₅ , X ₃₆ , X ₃₇ , X ₃₈ and X ₃₉ are absent.

53. The peptide of embodiment 49, wherein X ₃₂ , X ₃₃ , X ₃₄ , X ₃₅ , X _{36 ,} X _{37 ,} X ₃₈ and X ₃₉ are absent.

54. The peptide of embodiment 49, wherein X ₃₀ , X ₃₁ , X ₃₂ , X ₃₃ , X ₃₄ , X ₃₅ , X _{36 ,} X _{37 ,} X ₃₈ and X ₃₉ are absent.

55. The peptide according to embodiment 49 or 50, wherein X ₃₂ X ₃₃ X ₃₄ X ₃₅ is selected from the group of SSGA, ESGA and SKGA.

56. The peptide of embodiment 55, wherein X ₃₂ X ₃₃ X ₃₄ X ₃₅ is SSGA.

57. The peptide of embodiment 55, wherein X ₃₂ X ₃₃ X ₃₄ X ₃₅ is ESGA.

58. The peptide of embodiment 55, wherein X ₃₂ X ₃₃ X ₃₄ X ₃₅ is SKGA.

59. The peptide according to any one of embodiments 47 to 58, wherein the peptide has an amide modification at the C-terminus.

60. The amino acid sequence of embodiment 49, wherein the peptide is

YX ₂ EGTX ₆ TSDYSX ₁₂ X ₁₃ LEX ₁₆ QAAX ₂₀ X ₂₁ FX ₂₃ X ₂₄ WLLX ₂₈ GGPSSGAPPPS (SEQ ID NO: 37)

From here;

X ₂ is Aib or A,

X ₆ is F or V,

X ₁₂ is I or Y,

X ₁₃ is Y, A, L or I;

X ₁₆ is K or E,

X ₂₀ is Q, R, E, H,

X ₂₁ is A or E,

X ₂₃ is I or V,

X ₂₄ is E, Q or N,

X ₂₈ is A or R;

peptide.

61. The amino acid sequence of embodiment 48 of the peptide is

YX ₂ EGTX ₆ TSDYSX ₁₂ X ₁₃ LEX ₁₆ QAAX ₂₀ X ₂₁ FX ₂₃ X ₂₄ WLLX ₂₈ GGPESGAPPPS (SEQ ID NO: 38)

From here;

X ₂ is Aib or A,

X ₆ is F or V,

X ₁₂ is I or Y,

X ₁₃ is Y, A, L or I;

X ₁₆ is K or E,

X ₂₀ is Q, R, E, H,

X ₂₁ is A or E,

X ₂₃ is I or V,

X ₂₄ is E, Q or N,

X ₂₈ is A or R;

peptide.

62. The amino acid sequence of embodiment 49, wherein the peptide is

YX ₂ EGTX ₆ TSDYSX ₁₂ X ₁₃ LEX ₁₆ QAAX ₂₀ X ₂₁ FX ₂₃ X ₂₄ WLLX ₂₈ GGPSKGAPPPS (SEQ ID NO: 39)

From here;

X ₂ is Aib or A,

X ₆ is F or V,

X ₁₂ is I or Y,

X ₁₃ is Y, A, L or I;

X ₁₆ is K or E,

X ₂₀ is Q, R, E, H,

X ₂₁ is A or E,

X ₂₃ is I or V,

X ₂₄ is E, Q or N,

X ₂₈ is A or R;

peptide.

63. The peptide according to embodiment 49, wherein the amino acid sequence of the peptide is any one of SEQ ID NOs: 1-27 or 33-35.

64. The peptide according to embodiment 63, wherein the peptide has a C-terminal amide modification.

65. The peptide according to any one of embodiments 49 to 64, wherein X ₁₃ LEX ₁₆ QAAX ₂₀ X ₂₁ FX ₂₃ X ₂₄ is selected from the group consisting of LLEKQAAREFIN, LLEKQAAREFIE, LLEKQAAQEFIE and LLEEQAAREFIE.

66. The peptide according to any one of embodiments 49 to 64, wherein the peptide is human GLP-1 and GIP receptor in vitro with an EC ₅₀ of less than 20 pM when measured without HSA in a CRE luciferase reporter assay as described in example 2 activating the peptide.

67. The peptide according to any one of embodiments 49 to 64, wherein X ₁₆ is K.

68. The peptide according to any one of embodiments 49 to 64, wherein X ₃₃ is K.

69. The peptide according to embodiment 49, wherein the amino acid sequence of the peptide is any one of SEQ ID NOs: 15, 28, 29, 31, 32 or 43.

70. The peptide according to embodiment 49, wherein the amino acid sequence of the peptide is SEQ ID NO: 31.

71. A method for preparing a compound according to any one of embodiments 1 to 40.

72. A method for preparing a peptide according to any one of embodiments 49 to 64.

73. A method for preparing a peptide according to any one of embodiments 49 to 70.

Methods and Examples

abbreviation list

The following abbreviations, listed in alphabetical order, are used below:

Ac: Acetyl

Ado (also referred to as OEG): 8-amino-3,6-dioxaoctanoic acid

Aib: α-aminoisobutyric acid

API: Active Pharmaceutical Ingredients

AUC: Area under the curve

BG: blood sugar

BHK: Baby hamster kidney

Boc: tert -butyloxycarbonyl

BW: weight

CAS: Chemical Abstracts Service

Cl-HOBt: 6-chloro-1-hydroxybenzothiazole

DCM: dichloromethane

DIC: diisopropylcarbodiimide

DIPEA: N,N - diisopropylethylamine

DMEM: Dulbecco's Modied Eagle's Medium

DPBS: Dulbecco Phosphate Buffered Saline

EDTA: Ethylenediaminetetraacetic acid

ELISA: Enzyme-Linked Immunosorbent Assay

equiv: molar equivalent

FBS: Fetal Bovine Serum

Fmoc: 9-fluorenylmethyloxycarbonyl

GIP: Glucose-dependent Insulin Releasing Polypeptide

GIPR: Glucose-dependent Insulin Releasing Polypeptide Receptor

GLP-1: Glucagon-Like Peptide 1

GLP-1R: Glucagon-Like Peptide 1 Receptor

h: time

HEPES: 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid

HFIP: 1,1,1,3,3,3-hexafluoro-2-propanol or hexafluoroisopropanol

HPLC: High Performance Liquid Chromatography

HSA: Human Serum Albumin

i.p.: intraperitoneal

IPGTT: Intraperitoneal Glucose Tolerance Test

i.v.: intravenous

LCMS: Liquid Chromatography Mass Spectrometry

LYD: Landrace Yorkshire Duroc

MeCN: acetonitrile

MeOH: methanol

mM: millimoles

mmol: millimoles

min: minutes

Mtt: 4-methyltrityl

MW: molecular weight

nM: nanomolar

NMP: 1-methyl-pyrrolidin-2-one

OEG (also referred to as Ado): 8-amino-3,6-dioxaoctanoic acid

OtBu: tert -butyl ester

Oxyma Pure®: Cyano-hydroxyimino-acetic acid ethyl ester

Pbf: 2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl

PBS: Phosphate Buffered Saline

PD: Pharmacodynamics

PK: Pharmacokinetics

pM: picomolar

RP: inverse

RP-HPLC: Reversed Phase High Performance Liquid Chromatography

rpm: revolutions/min

RT: room temperature

Rt: retention time

s.c.: subcutaneously

SD: standard deviation

SEC-HPLC: Size Exclusion High Performance Liquid Chromatography

SEM: standard error of the mean

SNAC: Sodium N- [8 - (2-hydroxybenzoyl) amino] caprylate

SPPS: Solid-Phase Peptide Synthesis

tBu: tert -butyl

TFA: trifluoroacetic acid

TIS: triisopropylsilane

Trt: triphenylmethyl or trityl

Trx: tranexamic acid

general manufacturing method

Methods for solid-phase peptide synthesis (SPPS methods including methods for deprotection of amino acids, methods for cleaving and purifying peptides from resins) as well as methods for detecting and characterizing the resulting peptides (LCMS methods) are described below.

The resin used to prepare the C-terminal peptide amide was H-Rink Amide-ChemMatrix resin (eg, 0.5 mmol/g loading). Unless specifically stated otherwise, the Fmoc protected amino acid derivatives used are based on the recommended standards: Fmoc-Ala-OH, Fmoc-Arg(Pbf)-OH, Fmoc-Asn(Trt)-OH, Fmoc-Asp(OtBu). )-OH, Fmoc-Cys(Trt)-OH, Fmoc-Gln(Trt)-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Gly-OH, Fmoc-His(Trt)-OH, Fmoc-Ile- OH, Fmoc-Leu-OH, Fmoc-Lys(Boc)-OH, Fmoc-Met-OH, Fmoc-Phe-OH, Fmoc-Pro-OH, Fmoc-Ser(tBu)-OH, Fmoc-Thr(tBu) -OH, Fmoc-Trp(Boc)-OH, Fmoc-Tyr(tBu)-OH, Fmoc-Val-OH, Fmoc-Lys(Mtt)-OH, Fmoc-Aib-OH, etc. (e.g., AAPPTEC, Anaspec, Bachem, ChemImpex, Iris Biotech, Midwest Biotech, Gyros Protein Technologies). Unless otherwise specified, the natural L-form of the amino acid is used. Either use reagents with a pre-attached Boc group (e.g., Boc-Tyr(tBu)-OH for peptides with Tyr at the N-terminus), or attach an amino acid to the peptide N-terminus followed by attachment to the Boc protecting group. The N-terminal amino acid was Boc protected at the alpha amino group by exchanging the N-terminal Fmoc protecting group.

For modular albumin binding moieties using SPPS, the following appropriately protected building blocks such as, but not limited to, Fmoc-8-amino-3,6-dioxaoctanoic acid ( Fmoc-Ado-OH), Boc-Lys(Fmoc)-OH, Fmoc-Glu-OtBu, hexadecanedioic acid mono-tert-butyl ester, octadecanedioic acid mono-tert-butyl ester, nonadecanedioic acid mono- Tert-butyl ester, eicosandioic acid mono-tert-butyl ester, tetradecanedioic acid mono-tert-butyl ester, or 4-(9-carboxynonyloxy)benzoic acid tert-butyl ester were used. All operations mentioned below were performed within the range of 0.1-0.2 mmol synthesis scale.

1. Synthesis of Resin-Bonded Protected Peptide Backbone

Method: SPPS_A

SPPS was performed using Fmoc-based chemistry on a Protein Technologies SymphonyX solid-phase peptide synthesizer using a manufacturer-supplied protocol with minor modifications. Mixing was effected by occasional bubbling with nitrogen. Stepwise assembly was performed using the following steps: 1) pre-swelling of the resin in DMF; 2) Fmoc-deprotection step with the use of 20% (v/v) piperidine in DMF for 2 treatments of 10 min each; 3) washing with DMF to remove piperidine; 4) Binding of Fmoc-amino acids by addition of Fmoc-amino acids (12 equiv) and Oxyma Pure® (12 equiv) as 0.6 M solution in DMF, respectively, followed by addition of DIC (12 equiv) as 1.2 M solution in DMF followed by DMF was added to reduce the final concentration of each component to 0.3 M, followed by mixing for 0.5 to 4 hours; 4) washing with DMF to remove excess reagent; 5) Final cleaning with DCM upon completion of assembly. Some amino acids, such as, but not limited to, those following a sterically constrained amino acid (eg, Aib), were combined through extended reaction times (eg, 4 hours) to allow the reaction to complete.

Method: SPPS_B

Protected peptidyl resins were synthesized according to the Fmoc strategy on an Applied Biosystems 431A solid phase peptide synthesizer using the manufacturer-supplied generic Fmoc protocol. Mixing was performed by bubbling with nitrogen occasionally with a vortex. Stepwise assembly was performed using the following steps: 1) Activation of Fmoc-amino acid by dissolution of solid Fmoc-acid (10 equiv) in Cl-HOBt (10 equiv) as a 1 M solution in NMP followed by 1 in NMP Addition of DIC (10 equiv) as M solution followed by simultaneous mixing to steps 2 and 3; 2) Fmoc-deprotection with use of 20% (v/v) piperidine in NMP for one third treatment followed by a second treatment of 15 minutes; 3) washing with NMP to remove piperidine; 4) after adding the activated Fmoc-amino acid solution to the resin, mixing for 45 to 90 minutes; 4) washing with NMP to remove excess reagent; 5) Final cleaning with DCM upon completion of assembly. The standard protected amino acid derivatives listed above are supplied in a pre-weighed cartridge (eg, from Midwest Biotech) and the non-standard derivatives are manually weighed. Some amino acids, such as, but not limited to, those following a sterically constrained amino acid (eg, Aib), were "double bonded" to allow the reaction to complete, which occurred after the first binding (eg, 45 min). After that, the resin was drained, more reagents (Fmoc-amino acid, DIC, Cl-HOBt) were added, and the mixture was reacted again (eg 45 min).

Method: SPPS_C

SPPS was performed using Fmoc-based chemistry on a PreludeX solid-phase peptide synthesizer using a manufacturer-supplied protocol with minor modifications. Mixing was performed by shaking at 350 rpm and occasionally bubbling with nitrogen. Stepwise assembly was performed using the following steps: 1) pre-swelling of the resin in DMF; 2) Fmoc-deprotection step with the use of 20% (v/v) piperidine in DMF for one-third treatment at 70° C.; 3) washing with DMF to remove piperidine; 4) Binding of Fmoc-amino acids by addition of Fmoc-amino acids (12 equiv) and Oxyma Pure® (12 equiv) as a 0.4 M solution in DMF, respectively, followed by addition of DIC (12 equiv) as a 1.2 M solution in DMF, followed by mixing at 70° C. for 5 minutes; 4) washing with DMF to remove excess reagent; 5) Final cleaning with DCM upon completion of assembly. Some amino acids, such as, but not limited to, those following a sterically constrained amino acid (eg, Aib), were combined through extended reaction times (eg, 15 minutes) to allow the reaction to complete.

2. Attachment of Substituents to the Resin-Bonded Protected Peptide Backbone

Method: SC_A

The resin was washed with 30% HFIP in DCM for 2 treatments of 45 min each, followed by washing with DCM and DMF to remove the N-epsilon-lysine protected Mtt protecting group. On Protein Technologies SymphonyX solid phase peptide synthesizer, Boc-Lys(Fmoc)-OH, Fmoc-8-amino-3,6-dioxaoctanoic acid, Fmoc-Glu-OtBu, hexadecanedioic acid mono- tert -butyl ester, octadecane Acylation using the protocol described in Method SPPS_A, employing the stepwise addition of building blocks such as, but not limited to, dioic acid mono- tert -butyl ester, and eicosandioic acid mono- tert -butyl-ester was performed.

Method: SC_B

The resin was washed with 30% HFIP in DCM for 2 treatments of 45 min each, followed by washing with DCM and DMF to remove the N-epsilon-lysine protected Mtt protecting group. On an Applied Biosystems 431A solid phase peptide synthesizer, Boc-Lys(Fmoc)-OH, Fmoc-8-amino-3,6-dioxaoctanoic acid, Fmoc-Glu-OtBu, hexadecanedioic acid mono- tert -butyl ester, octadecane Acylation using the protocol described in Method SPPS_B, employing the stepwise addition of building blocks such as, but not limited to, dioic acid mono- tert -butyl ester, and eicosandioic acid mono- tert -butyl-ester was performed.

Method: SC_C

The resin was washed with 30% HFIP in DCM for 2 treatments of 45 min each, followed by washing with DCM and DMF to remove the N-epsilon-lysine protected Mtt protecting group. On Protein Technologies PreludeX solid phase peptide synthesizer, Boc-Lys(Fmoc)-OH, Fmoc-8-amino-3,6-dioxaoctanoic acid, Fmoc-Glu-OtBu, hexadecanedioic acid mono- tert -butyl ester, octadecane Acylation using the protocol described in Method SPPS_C, employing the stepwise addition of building blocks such as, but not limited to, dioic acid mono- tert -butyl ester, and eicosandioic acid mono- tert -butyl-ester. was performed.

3. Cleavage and purification of resin-bound peptides

Method: CP_A

After the side chain synthesis was completed, the peptidyl resin was washed with DCM and dried, treated with TFA/water/TIS (95:2.5:2.5 v/v/v) for about 2 hours, and then precipitated with diethyl ether. made it The precipitate was washed with diethyl ether, dissolved in a suitable solvent (eg 2:1 water/MeCN) and left until all labile adducts were resolved. Purification was performed using a Phenomenex Luna C8(2) column (10 μM particle size, 100 Å pore size, 250 x 21.2 mm dimensions) by reverse-phase preparative HPLC (Waters 2545 binary gradient module, Waters 2489 UV/Visible light detector, Waters Fraction Collector III). was performed with Impurity separation and elution of the product was achieved using an increasing gradient of MeCN in water containing 0.1% TFA. Analytical LCMS identified the relevant fractions and determined the purity. Fractions containing the pure desired product were pooled and lyophilized to give the TFA salt of the peptide as a white solid.

4. Salt exchange from TFA to sodium salt:

Method: SX_A:

The lyophilized peptide isolated from Method CP_A is dissolved in an appropriate aqueous buffer (e.g., 4:1 water/MeCN, 0.2 M sodium acetate) to 5-20 mg/mL and, if necessary, from pH 7 to pH 7 with 1 M NaOH. 8 to achieve complete solubility. The buffer containing the peptide was salt exchanged using a Sep-Pak C18 cartridge (0.5-2 g). The cartridge was first equilibrated using 4 column volumes of isopropanol followed by 4 column volumes of MeCN followed by 8 column volumes of water. The peptide solution was poured into the cartridge and the overflow was poured back to ensure complete retention of the peptide. Wash the cartridge with 4 column volumes of water and then 10 column volumes of buffer (eg, pH 7.5) containing a sodium salt such as, but not limited to, NaHCO ₃ , NaOAc, or Na ₂ HPO ₄ . did The column was washed with 4 column volumes of water and the peptides were eluted with 50-80% MeCN in 5-10 column volumes of water. Lyophilization of the peptide containing eluate gave the peptide sodium salt as a white solid, which was used as such.

Common detection and characterization methods

LCMS method:

Method: LCMS_A

LCMS_A was performed in an environment configured with an Agilent 1260 Infinity series UPLC system and an Agilent Technologies 6120 Quadrupole MS. Eluent: A: 0.05% TFA in water; B: 0.05% TFA in 9:1 MeCN/water.

Analysis was performed by injecting an appropriate volume of sample onto a column at room temperature (column temperature 37° C. and eluting with a gradient of A and B. Column: Phenomenex Kinetex C8, 2.6 μm, 100 Å, 4.6×75 mm. Gradient run time : Linear 10-80% over 10 min at a flow rate of 1.0 mL/min B. Detection: Diode Array Detector set to 214 nm MS Ionization Mode: API-ES, Positive Polarity MS Scan Mass Range: 500 to 2000 amu • The most abundant isotope of each m/z reported.

Method: LCMS_B

LCMS_B was performed in an environment configured with an Agilent 1260 Infinity series UPLC system and an Agilent Technologies 6120 Quadrupole MS. Eluent: A: 0.05% TFA in water; B: 0.05% TFA in 9:1 MeCN/water.

Analysis was performed by injecting an appropriate volume of sample onto a column at room temperature (column temperature 37° C. and eluting with a gradient of A and B. Column: Phenomenex Kinetex C8, 2.6 μm, 100 Å, 4.6×75 mm. Gradient run time : Linear 20-100% over 10 min at a flow rate of 1.0 mL/min B. Detection: Diode Array Detector set to 214 nm MS Ionization Mode: API-ES, Positive Polarity MS Scan Mass Range: 500 to 2000 amu • The most abundant isotope of each m/z reported.

While specific features of the invention have been illustrated and described herein, many modifications, substitutions, changes and equivalents will now occur to those skilled in the art. Accordingly, it is to be understood that the appended embodiments are intended to embrace all such modifications and variations that fall within the true spirit of the present invention.

Example 1: Synthesis of compounds

Compounds are described below using single letter amino acid codes, with the exception of Aib. A substituent is included after the lysine (K) residue to which it is attached.

compound 1

Y-Aib-EGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAQEFVNWLLAGGPSSGAPPPS-NH ₂

SEQ ID NO: 1 with a C-terminal amide modification; Substituent: C; Substituent Position: K16

Synthesis method: SPPS_A; SC_B; CP_A

Calculated molecular weight (average): 4873.5 Da

LCMS_A: Rt = 6.0 min; Found as [M+3H] ³⁺ 1625.4, [M+4H] ⁴⁺ 1219.1.

compound 2

Y-Aib-EGTFTSDYSYYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoyl) amino)butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAQEFVNWLLAGGPSSGAPPPS-NH ₂

SEQ ID NO: 2 with a C-terminal amide modification; Substituent: C; Substituent Position: K16

Synthesis method: SPPS_A; SC_B; CP_A

Calculated molecular weight (average): 4923.5 Da

LCMS_A: Rt = 6.0 min; Found as [M+3H] ³⁺ 1641.8, [M+4H] ⁴⁺ 1237.5.

compound 3

Y-Aib-EGTFTSDYSYYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAQEFVNWLLAGGPSSGAPPPS-NH ₂

SEQ ID NO: 2 with a C-terminal amide modification; Substituent: B; Substituent Position: K16

Synthesis method: SPPS_A; SC_B; CP_A

Calculated molecular weight (average): 4895.4 Da

LCMS_A: Rt = 5.7 min; Found as [M+3H] ³⁺ 1632.7, [M+4H] ⁴⁺ 1224.6.

compound 4

Y-Aib-EGTFTSDYSYYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAQAFVNWLLAGGPSSGAPPPS-NH ₂

SEQ ID NO: 3 with a C-terminal amide modification; Substituent: B; Substituent Position: K16

Synthesis method: SPPS_A; SC_B; CP_A

Calculated molecular weight (average): 4837.4 Da

LCMS_A: Rt = 5.7 min; Found as [M+3H] ³⁺ 1613.3, [M+4H] ⁴⁺ 1210.1.

compound 5

Y-Aib-EGTFTSDYSYYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAQAFVNWLLAGGPSSGAPPPS-OH

SEQ ID NO: 3; Substituent: B; Substituent Position: K16

Synthesis method: SPPS_B; SC_B; CP_A

Calculated molecular weight (average): 4838.4 Da

LCMS_A: Rt = 5.7 min; Found as [M+3H] ³⁺ 1613.6, [M+4H] ⁴⁺ 1210.4.

compound 6

Y-Aib-EGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFVNWLLAGGPSSGAPPPS-NH ₂

SEQ ID NO: 4 with a C-terminal amide modification; Substituent: B; Substituent Position: K16

Synthesis method: SPPS_B; SC_B; CP_A

Calculated molecular weight (average): 4873.5 Da

LCMS_A: Rt = 5.6 min; Found as [M+3H] ³⁺ 1625.3, [M+4H] ⁴⁺ 1219.1.

compound 7

Y-Aib-EGTFTSDYSIYLE-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl] amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-QAAREFVNWLLAGGPSSGAPPPS-NH ₂

SEQ ID NO: 4 with a C-terminal amide modification; Substituent: A; Substituent Position: K16

Synthesis method: SPPS_B; SC_B; CP_A

Calculated molecular weight (average): 4907.4 Da

LCMS_A: Rt = 6.3 min; Found as [M+3H] ³⁺ 1636.5, [M+4H] ⁴⁺ 1227.9.

compound 8

Y-Aib-EGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFINWLLAGGPSSGAPPPS-NH ₂

SEQ ID NO: 5 with a C-terminal amide modification; Substituent: B; Substituent Position: K16

Synthesis method: SPPS_A; SC_A; CP_A

Calculated molecular weight (average): 4887.5 Da

LCMS_A: Rt = 5.7 min; Found as [M+3H] ³⁺ 1629.8, [M+4H] ⁴⁺ 1222.8.

compound 9

Y-Aib-EGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFIEWLLAGGPSSGAPPPS-OH

SEQ ID NO: 6; Substituent: B; Substituent Position: K16

Synthesis method: SPPS_A; SC_B; CP_A

Calculated molecular weight (average): 4903.5 Da

LCMS_A: Rt = 5.8 min; Found as [M+3H] ³⁺ 1635.2, [M+4H] ⁴⁺ 1226.8.

compound 10

Y-Aib-EGTFTSDYSIYLE-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl] amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-QAAREFIEWLLAGGPSSGAPPPS-OH

SEQ ID NO: 6; Substituent: A; Substituent Position: K16

Synthesis method: SPPS_A; SC_B; CP_A

Calculated molecular weight (average): 4937.5 Da

LCMS_A: Rt = 6.5 min; Found as [M+3H] ³⁺ 1646.6, [M+4H] ⁴⁺ 1235.1.

compound 11

Y-Aib-EGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFINWLLAGGPSSGAPPPS-OH

SEQ ID NO: 5; Substituent: B; Substituent Position: K16

Synthesis method: SPPS_A; SC_C; CP_A

Calculated molecular weight (average): 4888.5 Da

LCMS_A: Rt = 5.7 min; Found as [M+3H] ³⁺ 1630.2, [M+4H] ⁴⁺ 1222.9.

compound 12

Y-Aib-EGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFIEWLLRGGPSSGAPPPS-OH

Synthesis method: SPPS_A; SC_C; CP_A

SEQ ID NO: 7; Substituent: B; Substituent Position: K16

Calculated molecular weight (average): 4988.6 Da

LCMS_A: Rt = 5.7 min; Found as [M+3H] ³⁺ 1663.6, [M+4H] ⁴⁺ 1248.0.

compound 13

Y-Aib-EGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFIEWLLRGGPESGAPPPS-OH

SEQ ID NO: 8; Substituent: B; Substituent Position: K16

Synthesis method: SPPS_A; SC_C; CP_A

Calculated molecular weight (average): 5030.6 Da

LCMS_A: Rt = 5.7 min; Found as [M+3H] ³⁺ 1677.7, [M+4H] ⁴⁺ 1258.4.

compound 14

Y-Aib-EGTFTSDYSIALE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFINWLLAGGPSSGAPPPS-OH

SEQ ID NO: 9; Substituent: B; Substituent Position: K16

Synthesis method: SPPS_A; SC_C; CP_A

Calculated molecular weight (average): 4796.4 Da

LCMS_A: Rt = 5.7 min; Found as [M+3H] ³⁺ 1599.6, [M+4H] ⁴⁺ 1199.8.

compound 15

Y-Aib-EGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFINWLLAGGPSSGAPPPS-OH

SEQ ID NO: 10; Substituent: B; Substituent Position: K16

Synthesis method: SPPS_A; SC_A; CP_A; SX_A

Calculated molecular weight (average): 4838.5 Da

LCMS_A: Rt = 5.8 min; Found as [M+3H] ³⁺ 1613.4, [M+4H] ⁴⁺ 1210.3.

compound 16

Y-Aib-EGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFINWLLAGGPSSGAPPPS-OH

SEQ ID NO: 5; Substituent: C; Substituent Position: K16

Synthesis method: SPPS_A; SC_A; CP_A

Calculated molecular weight (average): 4916.5 Da

LCMS_A: Rt = 6.0 min; Found as [M+3H] ³⁺ 1639.6, [M+4H] ⁴⁺ 1229.9.

compound 17

YAEGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl] amino]hexanoyl]amino]hexanoyl]-QAAREFINWLLAGGPSSGAPPPS-OH

compound 18

Y-Aib-EGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFINWLLAG-OH

SEQ ID NO: 12; Substituent: B; Substituent Position: K16

Synthesis method: SPPS_A; SC_A; CP_A

Calculated molecular weight (average): 4003.6 Da

LCMS_A: Rt = 6.2 min; Found as [M+3H] ³⁺ 1335.3, [M+4H] ⁴⁺ 1001.7.

compound 19

Y-Aib-EGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFINWLLAGGP-OH

SEQ ID NO: 13; Substituent: B; Substituent Position: K16

Synthesis method: SPPS_A; SC_A; CP_A

Calculated molecular weight (average): 4157.8 Da

LCMS_A: Rt = 6.1 min; Found as [M+3H] ³⁺ 1386.6, [M+4H] ⁴⁺ 1040.3.

compound 20

Y-Aib-EGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFINWLLAGGPSS-OH

SEQ ID NO: 14; Substituent: B; Substituent Position: K16

Synthesis method: SPPS_A; SC_A; CP_A

Calculated molecular weight (average): 4331.9 Da

LCMS_A: Rt = 5.9 min; Found as [M+3H] ³⁺ 1444.7, [M+4H] ⁴⁺ 1083.7.

compound 21

Y-Aib-EGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAR-EFINWLLAGGPSSGA-OH

SEQ ID NO: 15; Substituent: B; Substituent Position: K16

Synthesis method: SPPS_A; SC_A; CP_A

Calculated molecular weight (average): 4460.0 Da

LCMS_A: Rt = 5.9 min; Found as [M+3H] ³⁺ 1487.5, [M+4H] ⁴⁺ 1116.1.

compound 22

Y-Aib-EGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFVNWLLAGGPSSGAPPPS-OH

SEQ ID NO: 16; Substituent: B; Substituent Position: K16

Synthesis method: SPPS_A; SC_A; CP_A

Calculated molecular weight (average): 4824.4 Da

LCMS_A: Rt = 5.7 min; Found as [M+3H] ³⁺ 1608.9, [M+4H] ⁴⁺ 1206.9.

compound 23

Y-Aib-EGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFIQWLLAGGPSSGAPPPS-OH

SEQ ID NO: 17; Substituent: B; Substituent Position: K16

Synthesis method: SPPS_A; SC_A; CP_A

Calculated molecular weight (average): 4852.5 Da

LCMS_A: Rt = 5.9 min; Found as [M+3H] ³⁺ 1618.4, [M+4H] ⁴⁺ 1214.1.

compound 24

Y-Aib-EGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAQEFINWLLAGGPSSGAPPPS-OH

SEQ ID NO: 18; Substituent: B; Substituent Position: K16

Synthesis method: SPPS_A; SC_C; CP_A

Calculated molecular weight (average): 4810.4 Da

LCMS_A: Rt = 6.0 min; Found as [M+3H] ³⁺ 1604.0, [M+4H] ⁴⁺ 1203.4.

compound 25

Y-Aib-EGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAEEFINWLLAGGPSSGAPPPS-OH

SEQ ID NO: 19; Substituent: B; Substituent Position: K16

Synthesis method: SPPS_A; SC_C; CP_A

Calculated molecular weight (average): 4811.4 Da

LCMS_A: Rt = 6.1 min; Found as [M+3H] ³⁺ 1604.4, [M+4H] ⁴⁺ 1203.6.

compound 26

Y-Aib-EGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAHEFINWLLAGGPSSGAPPPS-OH

SEQ ID NO: 20; Substituent: B; Substituent Position: K16

Synthesis method: SPPS_A; SC_C; CP_A

Calculated molecular weight (average): 4819.4 Da

LCMS_A: Rt = 5.8 min; Found as [M+3H] ³⁺ 1607.1, [M+4H] ⁴⁺ 1205.5.

compound 27

Y-Aib-EGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFIEWLLRGGPSSGAPPPS-OH

SEQ ID NO: 21; Substituent: B; Substituent Position: K16

Synthesis method: SPPS_A; SC_C; CP_A

Calculated molecular weight (average): 4938.6 Da

LCMS_A: Rt = 5.7 min; Found as [M+3H] ³⁺ 1646.9, [M+4H] ⁴⁺ 1235.4.

compound 28

Y-Aib-EGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFIEWLLRGGPESGAPPPS-OH

SEQ ID NO: 22; Substituent: B; Substituent Position: K16

Synthesis method: SPPS_A; SC_A; CP_A; SX_A

Calculated molecular weight (average): 4980.6 Da

LCMS_A: Rt = 5.8 min; Found as [M+3H] ³⁺ 1660.7, [M+4H] ⁴⁺ 1246.0.

compound 29

Y-Aib-EGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAQEFIEWLLAGGPSSGAPPPS-OH

SEQ ID NO: 23; Substituent: B; Substituent Position: K16

Synthesis method: SPPS_A; SC_A; CP_A; SX_A

Calculated molecular weight (average): 4825.4 Da

LCMS_A: Rt = 6.1 min; Found as [M+3H] ³⁺ 1609.3, [M+4H] ⁴⁺ 1207.0.

compound 30

Y-Aib-EGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAHEFIEWLLAGGPSSGAPPPS-OH

SEQ ID NO: 24; Substituent: B; Substituent Position: K16

Synthesis method: SPPS_A; SC_A; CP_A

Calculated molecular weight (average): 4834.4 Da

LCMS_A: Rt = 5.8 min; Found as [M+3H] ³⁺ 1612.1, [M+4H] ⁴⁺ 1209.3.

compound 31

Y-Aib-EGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFIEWLLAGGPSSGAPPPS-OH

SEQ ID NO: 25; Substituent: B; Substituent Position: K16

Synthesis method: SPPS_A; SC_A; CP_A; SX_A

Calculated molecular weight (average): 4853.5 Da

LCMS_A: Rt = 5.9 min; Found as [M+3H] ³⁺ 1618.5, [M+4H] ⁴⁺ 1214.2.

compound 32

Y-Aib-EGTFTSDYSILLE-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl] amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-QAAREFIEWLLAGGPSSGAPPPS-OH

SEQ ID NO: 25; Substituent: A; Substituent Position: K16

Synthesis method: SPPS_A; SC_A; CP_A

Calculated molecular weight (average): 4887.4 Da

LCMS_A: Rt = 6.6 min; Found as [M+3H] ³⁺ 1629.8, [M+4H] ⁴⁺ 1222.8.

compound 33

Y-Aib-EGTVTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFINWLLAGGPSSGAPPPS-OH

SEQ ID NO: 26; Substituent: B; Substituent Position: K16

Synthesis method: SPPS_A; SC_A; CP_A

Calculated molecular weight (average): 4790.4 Da

LCMS_A: Rt = 5.8 min; Found as [M+3H] ³⁺ 1597.4, [M+4H] ⁴⁺ 1198.3.

compound 34

Y-Aib-EGTFTSDYSIILE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFINWLLAGGPSSGAPPPS-OH

SEQ ID NO: 27; Substituent: B; Substituent Position: K16

Synthesis method: SPPS_A; SC_A; CP_A

Calculated molecular weight (average): 4838.5 Da

LCMS_A: Rt = 5.8 min; Found as [M+3H] ³⁺ 1613.4, [M+4H] ⁴⁺ 1210.3.

compound 35

Y-Aib-EGTFTSDYSILLE-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl] amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-QAAREFINWLLAGGPSSGAPPPS-OH

SEQ ID NO: 10; Substituent: A; Substituent Position: K16

Synthesis method: SPPS_A; SC_A; CP_A

Calculated molecular weight (average): 4872.4 Da

LCMS_A: Rt = 6.6 min; Found as [M+3H] ³⁺ 1624.9, [M+4H] ⁴⁺ 1218.9.

compound 36

Y-Aib-EGTFTSDYSIILE-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl] amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-QAAREFINWLLAGGPSSGAPPPS-OH

SEQ ID NO: 27; Substituent: A; Substituent Position: K16

Synthesis method: SPPS_A; SC_A; CP_A

Calculated molecular weight (average): 4872.4 Da

LCMS_A: Rt = 6.5 min; Found as [M+3H] ³⁺ 1624.8, [M+4H] ⁴⁺ 1218.9.

compound 37

Y-Aib-EGTFTSDYSIYLEEQAAR-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-FINWLLAGGPSSGAPPPS-OH

SEQ ID NO: 28; Substituent: B; Substituent position: K21

Synthesis method: SPPS_A; SC_C; CP_A

Calculated molecular weight (average): 4888.5 Da

LCMS_A: Rt = 6.0 min; Found as [M+3H] ³⁺ 1630.3, [M+4H] ⁴⁺ 1223.0.

compound 38

Y-Aib-EGTFTSDYSIYLEE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-AAREFINWLLAGGPSSGAPPPS-OH

SEQ ID NO: 29; Substituent: B; Substituent Position: K17

Synthesis method: SPPS_C; SC_C; CP_A

Calculated molecular weight (average): 4889.5 Da

LCMS_A: Rt = 6.0 min; Found as [M+3H] ³⁺ 1630.6, [M+4H] ⁴⁺ 1223.2.

compound 39

Y-Aib-EGTFTSDYSILLEEQAAR-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-FINWLLAGGPSSGAPPPS-OH

SEQ ID NO: 30; Substituent: B; Substituent position: K21

Synthesis method: SPPS_A; SC_C; CP_A

Calculated molecular weight (average): 4838.5 Da

LCMS_A: Rt = 6.2 min; Found as [M+3H] ³⁺ 1613.5, [M+4H] ⁴⁺ 1210.4.

compound 40

Y-Aib-EGTFTSDYS-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-LLEEQAAREFIEWLLAGGPSSGAPPPS-OH

SEQ ID NO: 31; Substituent: B; Substituent Position: K12

Synthesis method: SPPS_A; SC_A; CP_A

Calculated molecular weight (average): 4869.4 Da

LCMS_A: Rt = 5.7 min; Found as [M+3H] ³⁺ 1623.8, [M+4H] ⁴⁺ 1218.2.

compound 41

Y-Aib-EGTFTSDYS-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-YLEEQAAREFINWLLAGGPSSGAPPPS-OH

SEQ ID NO: 32; Substituent: B; Substituent Position: K12

Synthesis method: SPPS_A; SC_A; CP_A

Calculated molecular weight (average): 4904.4 Da

LCMS_A: Rt = 5.7 min; Found as [M+3H] ³⁺ 1635.4, [M+4H] ⁴⁺ 1226.7.

compound 42

Y-Aib-EGTFTSDYSIYLEEQAAREFINWLLAGGPS-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-GAPPPS-OH

SEQ ID NO: 33; Substituent: B; Substituent Position: K33

Synthesis method: SPPS_A; SC_A; CP_A

Calculated molecular weight (average): 4930.5 Da

LCMS_A: Rt = 6.2 min; Found as [M+3H] ³⁺ 1644.2, [M+4H] ⁴⁺ 1233.4.

compound 43

Y-Aib-EGTFTSDYSILLEEQAAREFIEWLLAGGPS-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-GAPPPS-OH

SEQ ID NO: 34; Substituent: B; Substituent Position: K33

Synthesis method: SPPS_A; SC_A; CP_A

Calculated molecular weight (average): 4895.5 Da

LCMS_A: Rt = 6.3 min; Found as [M+3H] ³⁺ 1632.4, [M+4H] ⁴⁺ 1224.6.

compound 44

Y-Aib-EGTFTSDYSILLEEQAAREFINWLLAGGPS-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-GAPPPS-OH

SEQ ID NO: 35; Substituent: B; Substituent Position: K33

Synthesis method: SPPS_A; SC_A; CP_A

Calculated molecular weight (average): 4880.5 Da

LCMS_A: Rt = 6.2 min; Found as [M+3H] ³⁺ 1627.5, [M+4H] ⁴⁺ 1220.9.

compound 45

Y-Aib-EGTFTSDYSILLEEQAAREFIEWLLAGGPS-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl] amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-GAPPPS-OH

SEQ ID NO: 34; Substituent: A; Substituent Position: K33

Synthesis method: SPPS_A; SC_A; CP_A

Calculated molecular weight (average): 4929.5 Da

LCMS_A: Rt = 6.6 min; Found as [M+3H] ³⁺ 1643.8, [M+4H] ⁴⁺ 1233.1.

compound 46

Y-Aib-EGTFTSDYSILLEEQAAREFIEWLLAGGPS-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(15-carboxypentadecanoylamino) )butanoyl]amino]hexanoyl]amino]hexanoyl]-GAPPPS-OH

SEQ ID NO: 34; Substituent: E; Substituent Position: K33

Synthesis method: SPPS_A; SC_A; CP_A

Calculated molecular weight (average): 4867.5 Da

LCMS_A: Rt = 6.1 min; Found as [M+3H] ³⁺ 1623.1, [M+4H] ⁴⁺ 1217.6.

compound 47

Y-Aib-EGTFTSDYSILLEEQAAREFIEWLLAGGPS-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(15-carboxypentadecanoylamino)butanoyl] amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-GAPPPS-OH

SEQ ID NO: 34; Substituent: D; Substituent Position: K33

Synthesis method: SPPS_A; SC_A; CP_A

Calculated molecular weight (average): 4901.4 Da

LCMS_A: Rt = 6.3 min; Found as [M+3H] ³⁺ 1634.6, [M+4H] ⁴⁺ 1226.1.

Example 2: In vitro donor efficacy (CRE luciferase; whole cell)

The purpose of this example is to test the functional activity or efficacy of compounds in vitro at human GLP-1 and GIP receptors. In vitro functional potency is a measure of target receptor activation in whole-cell assays. The efficacy of the derivative of Example 1 was determined as described below. Human GLP-1 (7-37) and human GIP were included in the appropriate assays for comparison.

principle

In vitro functional efficacy was determined by measuring the response of the target receptor in a reporter gene assay in individual cell lines. Assays were performed on stably transfected BHK cell lines expressing one of the following G-protein coupled receptors, human GLP-1 receptors or human GIP receptors, each cell line expressing a cAMP response element (CRE) bound to a promoter. DNA for firefly and a gene for firefly luciferase (CRE luciferase). When each receptor is activated, cAMP is produced, and as a result, the luciferase protein is expressed. Upon completion of the assay incubation, a luciferase substrate (luciferin) is added, and the enzyme converts luciferin to oxyluciarin to produce bioluminescence. Luminescence is measured as a readout for the assay.

Cell Culture and Preparation

The cell line used in these assays is BHK cells with BHKTS13 as the parental cell line. The cell line was derived from a clone containing the CRE luciferase element and was established by further transfection with the respective human receptor to obtain the relevant cell line. BHK CRE luc2P hGLP-1R or BHK CRE luc2P hGIPR. Cells were cultured in a cell culture medium at 37° C. under 5% CO ₂ . They were aliquoted and stored in liquid nitrogen. Cells were maintained in continuous culture and seeded one day before each assay.

matter

The following chemicals were used in the assay: Pluronic 10% F-68 (Gibco 2404), Human Serum Albumin (HSA, Sigma A9511), 10% Fetal Bovine Serum (FBS, Invitrogen 16140-071), Egg White Orb Albumin (Sigma A5503), DMEM without phenol red (Gibco 21063-029), DMEM (Gibco 12430-054), 1 M Hepes (Gibco 15630), Glutamax 100x (Gibco 35050), G418 (Invitrogen 10131-027), he Gromycin (Invitrogen 10687-010), and steadylite plus (PerkinElmer 6016757).

buffer

GLP-1R cell culture medium consisted of DMEM medium with 10% FBS, 500 μg/mL G418, and 300 μg/mL hygromycin. GIPR cell culture medium consisted of DMEM medium with 10% FBS, 400 μg/mL G418, and 300 μg/mL hygromycin. Assay Buffer was DMEM without phenol red, 10 mM Hepes, 1 x Glutamax with 2x the final assay concentration of HSA. 1% Ovalbumin, and 0.1% Pluronic F-68. The assay buffer was mixed 1:1 with an equal volume of test compound in assay buffer to obtain a final assay concentration of HSA.

procedure

1) Cells were plated at 5000 cells/well and cultured overnight in assay plates.

2) Cells were washed once in DPBS.

3) Stocks of test compound and reference compound at concentrations ranging from 100 to 300 μM were diluted 1:150 in assay buffer. Compounds were then diluted 1:10 in column 1 of 96 deep well dilution plates and transported across rows to generate 3.5-fold, 12-point dilution curves.

4) Assay buffer (50 μl aliquot) with or without HSA was added to each well in the assay plate.

5) Transfer 50 μl of compound aliquot or blank from the dilution plate to the assay plate containing assay buffer with or without HSA.

6) The assay plate was incubated for 3 hours in an incubator at 37° C. under 5% CO ₂ .

7) Cells were washed once with DPBS.

8) An aliquot of 100 μl of DPBS was added to each well of the assay plate.

9) A 100 μl aliquot of Steadilite Plus reagent (photosensitive) was added to each well of the assay plate.

10) Each assay plate was covered with aluminum foil to block light, and shaken at room temperature at 250 RPM for 30 minutes.

11) Each assay plate was read with a microtiter plate reader.

Calculations and results

Data from the microtiter plate reader was first regressed in Excel to calculate x-axis, log scale concentrations based on stock concentrations of individual test compounds and dilutions of the assay. This data was then transferred to GraphPad Prism software for graphing and statistical analysis. The software performs a non-linear regression (log(agent) versus response). The EC ₅₀ values calculated by the software and recorded in pM are shown in Tables 1 to 3 below. At least two replicates were measured for each sample. The reported value is the average of the repeated values.

The compounds of the present invention exhibit potent functional activation of human GLP-1R and human GIPR receptors under given conditions.

Example 3: Pharmacokinetic study in minipigs

The purpose of this example is to determine the in vivo half-life of the derivatives of the present invention after intravenous (i.v.) administration to minipigs, ie, their prolonged residence time in the body and thereby their duration of action. This is done in pharmacokinetic (PK) studies, in which the terminal half-life of the corresponding derivative is determined. The terminal half-life usually refers to the time it takes for a specific plasma concentration to halve and is measured from the initial distribution phase.

research

Female Koettingen minipigs of approximately 7-14 months of age were obtained from Ellegaard Goettingen Minipigs (Dalmose, Denmark), and those weighing approximately 16-35 kg were used in the study. Minipigs were housed individually and fed limitedly once daily on the SDS minipig diet (Special Diets Services, Essex, UK).

After 3 weeks of acclimatization, each animal was implanted with two permanent central venous catheters in the back of the vena cava. After recovery of the animals for 1 week after surgery, they were used for repeated pharmacokinetic studies, and an appropriate wash-out period was placed in the middle of continuous administration of the derivatives.

Animals were not fed for about 18 hours prior to dosing and 0-4 hours after dosing, but were allowed to drink water ad libitum throughout the period.

The sodium salt of the compound of Example 1 was dissolved at a concentration of 20-40 nmol/mL in a buffer containing 0.007% polysorbate 20, 50 mM sodium phosphate, 70 mM sodium chloride, pH 7.4. The compound is injected intravenously via one catheter (generally in a volume corresponding to 1.5 to 2 nmol/kg, eg 0.1 ml/kg), at a predefined time point up to 14 days after administration (preferably the other blood was sampled through the catheter of Blood samples (eg, 0.8 mL) were pooled in 8 mM EDTA buffer and then centrifuged at 1942 G for 10 min at 4°C.

Sampling and Analysis

Plasma was pipetted into micronic tubes on dry ice and stored at -20°C until analysis of plasma concentrations of compounds using ELISA or similar antibody based assays or LCMS. Individual plasma concentration-time profiles were obtained from Phoenix WinNonLin ver. 6.4. (Pharsight Inc., Mountain View, CA, USA) was analyzed with a non-compartmental model, and the terminal half-life (harmonic mean) was determined.

result

The tested compounds of the present invention have very long half-lives compared to the half-lives of hGLP-1 and hGIP measured in humans, which are about 2-4 minutes and 5-7 minutes, respectively. Half-life measured in minipigs predicts a half-life in humans sufficient for at least once-weekly dosing via liquid injection or at least once-daily via oral tablets.

Example 5: Pharmacokinetic Study in Dogs

The purpose of this example is to determine the in vivo half-life and plasma exposure of a compound of the present invention after oral (p.o.) administration to beagle dogs, i.e., the final half-life and concentration of a test substance that reaches circulation over time. This was done in a pharmacokinetic (PK) study, in which these parameters of the compound were determined. The terminal half-life usually refers to the time it takes for a specific plasma concentration to halve and is measured from the initial distribution phase.

Preparation of tablet compositions

A tablet composition comprising a test substance and SNAC (sodium N- (8-(2-hydroxybenzoyl)amino)caprylate) can be prepared by roller compacting the test substance, as described, for example, in WO 2019/149880. Prepared according to methods known to those skilled in the art by mixing with SNAC and magnesium stearate. The amount of SNAC in the tablet composition was 100 to 300 mg, the amount of magnesium stearate in the tablet composition was 7.7 mg, and the target amount of each test substance in the tablet composition was 3 to 4 mg.

Animals, Dosing and Sampling

For the duration of the study, male beagle dogs aged 1 to 7 years and weighing 9 to 17 kg were included in the study. It was administered to dogs in a fasting state. Dogs are housed in packs (12 hours of light: 12 hours of shade) and fed individually, but are fed with medium-sized dog food (Royal Canin Medium Adult dog food, Royal Canin Products, China Branch, or Brogaarden A/S, Denmark) 1 It was restricted to feeding once a day. Dogs were used for repeated pharmacokinetic studies, with an appropriate washout period between each administration. An adequate acclimatization period was given prior to initiation of the primary pharmacokinetic study. All handling, dosing and blood collection of animals was performed by trained and skilled personnel. Dogs were fasted overnight before the study and 0-4 hours after dosing. Dogs were not allowed to drink water from 1 hour before dosing until 4 hours after dosing, but were otherwise allowed to drink water at any time during the entire period.

The composition was administered by oral administration to the dogs in groups of 6 to 8 dogs once. The tablets were administered in the following manner: 10 minutes prior to tablet administration, approximately 3 nmol/kg of SEQ ID NO: 48 was administered subcutaneously to the dog, and then the tablet was placed in the back of the dog's mouth to prevent chewing. Then, the dog's snout was closed and 10 mL of tap water was injected by syringe or gavage to facilitate swallowing of the tablet.

In order to adequately cover the total plasma concentration-time absorption profile of the test substance, one blood sample is taken prior to dosing, and 5, 10, 15, 20, 30 and 5, 10, 15, 20, 30 and Additional samples were taken after 45 minutes and after 1, 1.5, 2, 4, 7, 10, 24, 48, 72, 120, 144, 168, 192, 216 hours. For each blood sampling time point, approximately 0.8 mL of whole blood was drawn into an EDTA-coated 1.5 mL tube, and the tube was gently rotated to ensure that the sample was well mixed with the EDTA. Blood samples were pooled in EDTA buffer (8 mM) and then centrifuged at 2000 G for 10 minutes at 4°C. Plasma was pipetted into micronic tubes on dry ice and maintained at a temperature of -20°C or lower until analysis. Blood samples are taken as appropriate, e.g., through a venflon inserted into the cephalic vein of the forelimb for the first 2 hours, and then by syringe from the jugular vein for the remainder of the time points. Drain the first few drops from Benflon to prevent Benflon's heparinized saline from entering the sample). The first few drops were flushed from Benflon to prevent Benflon's heparinized saline from entering the sample.

All blood samples were collected in test tubes containing EDTA for stabilization and placed on ice until centrifugation. Plasma was separated from whole blood by centrifugation and stored at -20°C or lower until analysis.

analysis and calculation

Plasma was analyzed for the test substance using LC-MS (liquid-phase chromatography-mass spectrometry) known to those skilled in the art. The system is either a 10-valve interface module TurboFlow system, a CTC HTS PAL autosampler, an Accela 1250 pump, and a Thermo Fisher QExactive mass spectrometer equipped with a Hot Pocket column oven, or a valve interface module TurboFlow system, TriPlus RSI autosampler, Dionex UltiMate 3000 It consisted of a Thermo Fisher QExactive Plus mass spectrometer equipped with a pump and a Hot Pocket column oven. RP-HPLC separation was performed on a Phenomenex Onyx Monolitric C18 column (50 x 2.0 mm) at 30 °C at a flow rate of 0.8 mL/min, or an Agilent Poroshell 120 SB-C18 column (50 x 2.1 mm) at 60 °C at a flow rate of 0.4 mL/min. , 2.7 μm) using a linear gradient of 1:1 acetonitrile/methanol in 1% aqueous formic acid. The mass spectrometer was operated in either the cationization SIM mode or the cationization PRM mode.

For each individual animal, Pharsight Phoenix WinNonLin ver. for PK analysis. 6.4 Plasma concentration-time profiles were analyzed in a non-compartmental model in software or other related software, resulting in final half-life (t _1/2 ), maximum plasma concentration per dose (C _max /D), and time to maximum plasma concentration ( t _max ), and the area under the curve to infinity (AUC/D) per dose were determined. Summary statistics of pharmacokinetic results are presented as median (t _max ), harmonic mean (t _1/2 ), or arithmetic mean (C _max , AUC).

result

The tested compounds of the present invention demonstrate oral bioavailability in this model, as concentrations of the compounds in plasma are detected after oral administration (C _max /D > 0 and AUC/D > 0). In addition, the tested compounds of the present invention further have very long half-lives compared to the half-lives of hGLP-1 and hGIP measured in humans, which are about 2-4 minutes and 5-7 minutes, respectively.

While specific features of the invention have been illustrated and described herein, many modifications, substitutions, changes and equivalents will now occur to those skilled in the art. Accordingly, it is to be understood that the appended embodiments are intended to embrace all such modifications and variations that fall within the true spirit of the present invention.

SEQUENCE LISTING <110> Novo Nordisk A/S <120> CO-AGONISTS AT GLP-1 AND GIP RECEPTORS SUITABLE FOR ORAL DELIVERY <130> 200018 <160> 48 <170> PatentIn version 3.5 <210> 1 <211> 39 < 212> PRT <213> Artificial Sequence <220> <223> Synthetic <220> <221> MOD_RES <222> (2)..(2) <223> Aib <400> 1 Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Tyr Leu Glu Lys 1 5 10 15 Gln Ala Ala Gln Glu Phe Val Asn Trp Leu Leu Ala Gly Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Pro Ser 35 <210> 2 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <220> <221> MOD_RES <222> (2)..(2) <223> Aib <400> 2 Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Tyr Tyr Leu Glu Lys 1 5 10 15 Gln Ala Ala Gln Glu Phe Val Asn Trp Leu Leu Ala Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Ser 35 <210> 3 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <220> <221> MOD_RES <222> (2)..(2) <223> Aib <400> 3 Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Tyr Tyr Leu Glu Lys 1 5 10 15 Gln Ala Ala Gln Ala Phe Val Asn Trp Leu Leu Ala Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Pro Ser 35 <210> 4 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <220> <221> MOD_RES <222> (2)..(2) <223> Aib <400> 4 Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Tyr Leu Glu Lys 1 5 10 15 Gln Ala Ala Arg Glu Phe Val Asn Trp Leu Leu Ala Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Pro Ser 35 <210> 5 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <220> <221> MOD_RES <222> (2)..(2) <223> Aib <400> 5 Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Tyr Leu Glu Lys 1 5 10 15 Gln Ala Ala Arg Glu Phe Ile Asn Trp Leu Leu Ala Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Pro Ser 35 <210> 6 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <220 > <221> MOD_RES <222> (2)..(2) <223> Aib <400> 6 Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Tyr Leu Glu Lys 1 5 10 15 Gln Ala Ala Arg Glu Phe Ile Glu Trp Leu Leu Ala Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Pro Ser 35 <210> 7 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <220> <221 > MOD_RES <222> (2)..(2) <223> Aib <400> 7 Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Tyr Leu Glu Lys 1 5 10 15 Gln Ala Ala Arg Glu Phe Ile Glu Trp Leu Leu Arg Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Pro Ser 35 <210> 8 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <220> <221> MOD_RES < 222> (2)..(2) <223> Aib <400> 8 Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Tyr Leu Glu Lys 1 5 10 15 Gln Ala Ala Arg Glu Phe Ile Glu Trp Leu Leu Arg Gly Gly Pro Glu 20 25 30 Ser Gly Ala Pro Pro Pro Ser 35 <210> 9 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <220> <221> MOD_RES <222> ( 2)..(2) <223> Aib <400> 9 Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Ala Leu Glu Lys 1 5 10 15 Gln Ala Ala Arg Glu Phe Ile Asn Trp Leu Leu Ala Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Pro Ser 35 <210> 10 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <220> <221> MOD_RES <222> (2).. (2) <223> Aib <400> 10 Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Leu Leu Glu Lys 1 5 10 15 Gln Ala Ala Arg Glu Phe Ile Asn Trp Leu Leu Ala Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Ser 35 <210> 11 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 11 Tyr Ala Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Tyr Leu Glu Lys 1 5 10 15 Gln Ala Ala Arg Glu Phe Ile Asn Trp Leu Leu Ala Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Pro Ser 35 <210> 12 <211> 29 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <220> <221> MOD_RES <222> (2)..(2) <223> Aib <400> 12 Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Leu Leu Glu Lys 1 5 10 15 Gln Ala Ala Arg Glu Phe Ile Asn Trp Leu Leu Ala Gly 20 25 <210> 13 <211> 31 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <220> <2 21> MOD_RES <222> (2)..(2) <223> Aib <400> 13 Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Leu Leu Glu Lys 1 5 10 15 Gln Ala Ala Arg Glu Phe Ile Asn Trp Leu Leu Ala Gly Gly Pro 20 25 30 <210> 14 <211> 33 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <220> <221> MOD_RES <222> (2)..( 2) <223> Aib <400> 14 Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Leu Leu Glu Lys 1 5 10 15 Gln Ala Ala Arg Glu Phe Ile Asn Trp Leu Leu Ala Gly Gly Pro Ser 20 25 30 Ser <210> 15 <211> 35 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <220> <221> MOD_RES <222> (2)..(2) <223> Aib <400> 15 Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Leu Leu Glu Lys 1 5 10 15 Gln Ala Ala Arg Glu Phe Ile Asn Trp Leu Leu Ala Gly Gly Pro Ser 20 25 30 Ser Gly Ala 35 <210> 16 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <220> <221> MOD_RES <222> (2)..(2) <223> Aib <400> 16 Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Leu Leu Glu Lys 1 5 10 15 Gln Ala Ala Arg Glu Phe Val Asn Trp Leu Leu Ala Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Pro Ser 35 <210> 17 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <220> <221> MOD_RES <222> (2)..(2) <223> Aib <400> 17 Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Leu Leu Glu Lys 1 5 10 15 Gln Ala Ala Arg Glu Phe Ile Gln Trp Leu Leu Ala Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Pro Ser 35 <210> 18 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <220> <221> MOD_RES <222> (2)..(2) <223> Aib <400> 18 Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Leu Leu Glu Lys 1 5 10 15 Gln Ala Ala Gln Glu Phe Ile Asn Trp Leu Leu Ala Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Pro Ser 35 <210> 19 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <220 > <221> MOD_RES <222> (2)..(2) <223> Aib <400> 19 Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Leu Leu Glu Lys 1 5 10 15 Gln Ala Ala Glu Glu Phe Ile Asn Trp Leu Leu Ala Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Pro Ser 35 <210> 20 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <220 > <221> MOD_RES <222> (2)..(2) <223> Aib <400> 20 Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Leu Leu Glu Lys 1 5 10 15 Gln Ala Ala His Glu Phe Ile Asn Trp Leu Leu Ala Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Pro Ser 35 <210> 21 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <220> <221 > MOD_RES <222> (2)..(2) <223> Aib <400> 21 Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Leu Leu Glu Lys 1 5 10 15 Gln Ala Ala Arg Glu Phe Ile Glu Trp Leu Leu Arg Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Pro Ser 35 <210> 22 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <220> <221> MOD_RES < 222> (2)..(2) <223> Aib <400> 22 Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Leu Leu Glu Lys 1 5 10 15 Gln Ala Ala Arg Glu Phe Ile Glu Trp Leu Leu Ar g Gly Gly Pro Glu 20 25 30 Ser Gly Ala Pro Pro Ser 35 <210> 23 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <220> <221> MOD_RES <222> (2)..(2) <223> Aib <400> 23 Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Leu Leu Glu Lys 1 5 10 15 Gln Ala Ala Gln Glu Phe Ile Glu Trp Leu Leu Ala Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Pro Ser 35 <210> 24 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <220> <221> MOD_RES <222> (2) ..(2) <223> Aib <400> 24 Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Leu Leu Glu Lys 1 5 10 15 Gln Ala Ala His Glu Phe Ile Glu Trp Leu Leu Ala Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Pro Ser 35 <210> 25 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <220> <221> MOD_RES <222> (2)..( 2) <223> Aib <400> 25 Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Leu Leu Glu Lys 1 5 10 15 Gln Ala Ala Arg Glu Phe Ile Glu Trp Leu Leu Ala Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Pro Ser 35 <210> 26 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <220> <221> MOD_RES <222> (2)..(2) <223 > Aib <400> 26 Tyr Xaa Glu Gly Thr Val Thr Ser Asp Tyr Ser Ile Leu Leu Glu Lys 1 5 10 15 Gln Ala Ala Arg Glu Phe Ile Asn Trp Leu Leu Ala Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Pro Ser 35 <210> 27 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <220> <221> MOD_RES <222> (2)..(2) <223> Aib < 400> 27 Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Ile Leu Glu Lys 1 5 10 15 Gln Ala Ala Arg Glu Phe Ile Asn Trp Leu Leu Ala Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Ser 35 <210> 28 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <220> <221> MOD_RES <222> (2)..(2) <223> Aib <400> 28 Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Tyr Leu Glu Glu 1 5 10 15 Gln Ala Ala Arg Lys Phe Ile Asn Trp Leu Leu Ala Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Ser 35 <210> 29 < 211> 39 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <220> <221> MOD_RES <222> (2)..(2) <223> Aib <400> 29 Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Tyr Leu Glu Glu 1 5 10 15 Lys Ala Ala Arg Glu Phe Ile Asn Trp Leu Leu Ala Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Pro Ser 35 <210> 30 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <220> <221> MOD_RES <222> (2)..(2) <223> Aib <400> 30 Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Leu Leu Glu Glu 1 5 10 15 Gln Ala Ala Arg Lys Phe Ile Asn Trp Leu Leu Ala Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Ser 35 <210> 31 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <220> <221> MOD_RES <222> (2)..(2) <223> Aib <400> 31 Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Lys Leu Leu Glu Glu 1 5 10 15 Gln Ala Ala Arg Glu Phe Ile Glu Trp Leu Leu Ala Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Pro Ser 35 <210> 32 <211> 39 <212> PRT <213 > Artificial Sequence <220> <223> Synthetic <220> <221> MOD_RES <222> (2)..(2) <223> Aib <400> 32 Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Glu Glu 1 5 10 15 Gln Ala Ala Arg Glu Phe Ile Asn Trp Leu Leu Ala Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Pro Ser 35 <210> 33 <211> 39 <212> PRT <213> Artificial Sequence <220 > <223> Synthetic <220> <221> MOD_RES <222> (2)..(2) <223> Aib <400> 33 Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Tyr Leu Glu Glu 1 5 10 15 Gln Ala Ala Arg Glu Phe Ile Asn Trp Leu Leu Ala Gly Gly Pro Ser 20 25 30 Lys Gly Ala Pro Pro Pro Ser 35 <210> 34 <211> 39 <212> PRT <213> Artificial Sequence <220> <223 > Synthetic <220> <221> MOD_RES <222> (2)..(2) <223> Aib <400> 34 Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Leu Leu Glu Glu 1 5 10 15 Gln Ala Ala Arg Glu Phe Ile Glu Trp Leu Leu Ala Gly Gly Pro Ser 20 25 30 Lys Gly Ala Pro Pro Pro Ser 35 <210> 35 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Synthetic < 2 20> <221> MOD_RES <222> (2)..(2) <223> Aib <400> 35 Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Leu Leu Glu Glu 1 5 10 15 Gln Ala Ala Arg Glu Phe Ile Asn Trp Leu Leu Ala Gly Gly Pro Ser 20 25 30 Lys Gly Ala Pro Pro Pro Ser 35 <210> 36 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <220> < 221> MISC_FEATURE <222> (2)..(2) <223> Xaa is Aib or Ala <220> <221> MISC_FEATURE <222> (6)..(6) <223> Xaa is Phe or Val <220 > <221> MISC_FEATURE <222> (12)..(12) <223> Xaa is Ile or Tyr <220> <221> MISC_FEATURE <222> (13)..(13) <223> Xaa is Tyr, Ala , Leu or Ile <220> <221> MISC_FEATURE <222> (16)..(16) <223> Xaa is Lys or Glu <220> <221> MISC_FEATURE <222> (20)..(20) <223 > Xaa is Gln, Arg, Glu or His <220> <221> MISC_FEATURE <222> (21)..(21) <223> Xaa is Ala or Glu <220> <221> MISC_FEATURE <222> (23). .(23) <223> Xaa is Ile or Val <220> <221> MISC_FEATURE <222> (24)..(24) <223> Xaa is Glu, Gln or Asn <220> <221> MISC_FEATURE <222> (28)..(28) <223> Xaa is Ala or Arg <220> <221> MISC_FEATURE <222> (32)..(32) <223> Xaa is Glu, Ser or absent <220> <221> MISC_FEATURE <222> (33)..(33) <223> Xaa is Ser, Lys or absent <220> <221> MISC_FEATURE <222> (34)..(34) <223> Xaa is Gly or absent <220> <221> MISC_FEATURE <222> (35)..(35) <223> Xaa is Ala or absent <220> <221> MISC_FEATURE <222> (36)..(36) <223> Xaa is Pro or absent <220> <221> MISC_FEATURE <222> (37)..( 37) <223> Xaa is Pro or absent <220> <221> MISC_FEATURE <222> (38)..(38) <223> Xaa is Pro or absent <220> <221> MISC_FEATURE <222> (39). .(39) <223> Xaa is Ser or absent <400> 36 Tyr Xaa Glu Gly Thr Xaa Thr Ser Asp Tyr Ser Xaa Xaa Leu Glu Xaa 1 5 10 15 Gln Ala Ala Xaa Xaa Phe Xaa Xaa Trp Leu Leu Xaa Gly Gly Pro Xaa 20 25 30 Xaa Xaa Xaa Xaa Xaa Xaa Xaa 35 <210> 37 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <220> <221> MISC_FEATURE <222> (2) ..(2) <223> Xaa is Aib or Ala <220> <221> MISC_FEATURE <222> (6)..(6) <223> Xaa is Phe or Val <220> <221> MISC_FEATURE <222> ( 12)..(12) <223> Xaa is Ile or Tyr <220> <221> MISC_FEATURE <222> (13)..(13) <223> Xaa is Tyr, Ala, Leu or Ile <220> <221 > MISC_FEATURE <222> (16)..(16) <223> Xaa is Lys or Glu <220> <221> MISC_FEATURE <222> (20)..(20) <223> Xaa is Gln, Arg, Glu or His <220> <221> MISC_FEATURE <222> (21)..(21) <223> Xaa is Ala or Glu <220> <221> MISC_FEATURE <222> (23)..(23) <223> Xaa is Ile or Val <220> <221> MISC_FEATURE <222> (24)..(24) <223> Xaa is Glu, Gln or Asn <220> <221> MISC_FEATURE <222> (28)..(28) < 223> Xaa is Ala or Arg <400> 37 Tyr Xaa Glu Gly Thr Xaa Thr Ser Asp Tyr Ser Xaa Xaa Leu Glu Xaa 1 5 10 15 Gln Ala Ala Xaa Xaa Phe Xaa Xaa Trp Leu Leu Xaa Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Ser 35 <210> 38 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <220> <221> MISC_FEATURE <222> (2)..(2) <223> Xaa is Aib or Ala <220> <221> MISC_FEATURE <222> (6)..(6) <223> Xaa is Phe or Val <220> <221> MISC_FEATURE <222> (12)..( 12) <223> Xaa is Ile or Tyr <220> <2 21> MISC_FEATURE <222> (13)..(13) <223> Xaa is Tyr, Ala, Leu or Ile <220> <221> MISC_FEATURE <222> (16)..(16) <223> Xaa is Lys or Glu <220> <221> MISC_FEATURE <222> (20)..(20) <223> Xaa is Gln, Arg, Glu or His <220> <221> MISC_FEATURE <222> (21)..(21) <223> Xaa is Ala or Glu <220> <221> MISC_FEATURE <222> (23)..(23) <223> Xaa is Ile or Val <220> <221> MISC_FEATURE <222> (24)..( 24) <223> Xaa is Glu, Gln or Asn <220> <221> MISC_FEATURE <222> (28)..(28) <223> Xaa is Ala or Arg <400> 38 Tyr Xaa Glu Gly Thr Xaa Thr Ser Asp Tyr Ser Xaa Xaa Leu Glu Xaa 1 5 10 15 Gln Ala Ala Xaa Xaa Phe Xaa Xaa Trp Leu Leu Xaa Gly Gly Pro Glu 20 25 30 Ser Gly Ala Pro Pro Pro Ser 35 <210> 39 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <220> <221> MISC_FEATURE <222> (2)..(2) <223> Xaa is Aib or Ala <220> <221> MISC_FEATURE <222> ( 6)..(6) <223> Xaa is Phe or Val <220> <221> MISC_FEATURE <222> (12)..(12) <223> Xaa is Ile or Tyr <220> <221> MISC_FEATURE <222 > (13)..(13) <223> Xa a is Tyr, Ala, Leu or Ile <220> <221> MISC_FEATURE <222> (16)..(16) <223> Xaa is Lys or Glu <220> <221> MISC_FEATURE <222> (20).. (20) <223> Xaa is Gln, Arg, Glu or His <220> <221> MISC_FEATURE <222> (21)..(21) <223> Xaa is Ala or Glu <220> <221> MISC_FEATURE <222 > (23)..(23) <223> Xaa is Ile or Val <220> <221> MISC_FEATURE <222> (24)..(24) <223> Xaa is Glu, Gln or Asn <220> <221 > MISC_FEATURE <222> (28)..(28) <223> Xaa is Ala or Arg <400> 39 Tyr Xaa Glu Gly Thr Xaa Thr Ser Asp Tyr Ser Xaa Xaa Leu Glu Xaa 1 5 10 15 Gln Ala Ala Xaa Xaa Phe Xaa Xaa Trp Leu Leu Xaa Gly Gly Pro Ser 20 25 30 Lys Gly Ala Pro Pro Pro Ser 35 <210> 40 <211> 4 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 40 Ser Ser Gly Ala 1 <210> 41 <211> 4 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 41 Glu Ser Gly Ala 1 <210> 42 <211> 4 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 42 Ser Lys Gly Ala 1 <210> 43 <211> 12 <212> PRT <213> Artificial Sequence <2 20> <223> Synthetic <400> 43 Leu Leu Glu Lys Gln Ala Ala Arg Glu Phe Ile Asn 1 5 10 <210> 44 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Synthetic < 400> 44 Leu Leu Glu Lys Gln Ala Ala Arg Glu Phe Ile Glu 1 5 10 <210> 45 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 45 Leu Leu Glu Lys Gln Ala Ala Gln Glu Phe Ile Glu 1 5 10 <210> 46 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 46 Leu Leu Glu Glu Gln Ala Ala Arg Glu Phe Ile Glu 1 5 10 <210> 47 <211> 39 <212> PRT <213 > Artificial Sequence <220> <223> Synthetic <220> <221> MOD_RES <222> (2)..(2) <223> Aib <220> <221> MISC_FEATURE <222> (2)..(2) <223> Xaa is Aib or Ala <220> <221> MISC_FEATURE <222> (6)..(6) <223> Xaa is Phe or Val <220> <221> MISC_FEATURE <222> (12)..( 12) <223> Xaa is Ile or Tyr <220> <221> MISC_FEATURE <222> (13)..(13) <223> Xaa is Tyr, Ala, Leu or Ile <220> <221> MISC_FEATURE <222> (16)..(16) <223> Xaa is Lys or Glu <220> <221> MISC_FEATURE <222> (20)..(20) <223> Xaa is Gln, Arg, Glu or His <220> < 221> MISC_FEATURE <222> (21)..(21) <223> Xaa is Ala or Glu <220> <221> MISC_FEATURE <222> (23)..(23) <223> Xaa is Ile or Val <220 > <221> MISC_FEATURE <2 22> (24)..(24) <223> Xaa is Glu, Gln or Asn <220> <221> MISC_FEATURE <222> (28)..(28) <223> Xaa is Ala or Arg <220> < 221> MISC_FEATURE <222> (30)..(30) <223> Xaa is Gly or absent <220> <221> MISC_FEATURE <222> (31)..(31) <223> Xaa is Pro or absent <220 > <221> MISC_FEATURE <222> (32)..(32) <223> Xaa is Glu, Ser or absent <220> <221> MISC_FEATURE <222> (33)..(33) <223> Xaa is Ser , Lys or absent <220> <221> MISC_FEATURE <222> (34)..(34) <223> Xaa is Gly or absent <220> <221> MISC_FEATURE <222> (35)..(35) <223 > Xaa is Ala or absent <220> <221> MISC_FEATURE <222> (36)..(36) <223> Xaa is Pro or absent <220> <221> MISC_FEATURE <222> (37)..(37) <223> Xaa is Pro or absent <220> <221> MISC_FEATURE <222> (38)..(38) <223> Xaa is Pro or absent <220> <221> MISC_FEATURE <222> (39)..( 39) <223> Xaa is Ser or absent <400> 47 Tyr Xaa Glu Gly Thr Xaa Thr Ser Asp Tyr Ser Xaa Xaa Leu Glu Xaa 1 5 10 15 Gln Ala Ala Xaa Xaa Phe Xaa Xaa Trp Leu Leu Xaa Gly Xaa Xaa Xaa 20 25 30 Xaa Xaa Xaa Xaa Xaa Xaa Xaa 35 <210> 48 <211> 29 <212> PRT <213> Homo sapiens <400> 48 His Ser Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Ser 1 5 10 15Arg Arg Ala Gln Asp Phe Val Gln Trp Leu Met Asn Thr 20 25

Claims (15)

A compound comprising a peptide and a substituent or a pharmaceutically acceptable salt thereof, wherein the amino acid sequence of the peptide is as follows,
YX ₂ EGTX ₆ TSDYSX ₁₂ X ₁₃ LEX ₁₆ QAAX ₂₀ X ₂₁ FX ₂₃ X ₂₄ WLLX ₂₈ GX ₃₀ X ₃₁ X ₃₂ X ₃₃ X ₃₄ X ₃₅ X ₃₆ X ₃₇ X ₃₈ X ₃₉ (SEQ ID NO: 47)
It has an optional amide modification at the C-terminus,
From here;
X ₂ is Aib or A,
X ₆ is F or V,
X ₁₂ is I or Y,
X ₁₃ is Y, A, L or I;
X ₁₆ is K or E,
X ₂₀ is Q, R, E, H,
X ₂₁ is A or E,
X ₂₃ is I or V,
X ₂₄ is E, Q or N,
X ₂₈ is A or R,
X ₃₀ is G or absent;
X ₃₁ is P or absent,
X ₃₂ is E, S or absent,
X ₃₃ is S, K or absent,
X ₃₄ is G or absent;
X ₃₅ is A or absent,
X ₃₆ is P or absent;
X ₃₇ is P or absent;
X ₃₈ is P or absent;
X ₃₉ is S or absent;
wherein said substituent is attached to the peptide via a lysine (K) at position 16 or 33;
A compound or a pharmaceutically acceptable salt thereof. The compound of claim 1 , wherein X ₃₂ X ₃₃ X ₃₄ X ₃₅ is selected from the group of SSGA, ESGA and SKGA. According to claim 1, wherein the amino acid sequence of the peptide is as follows,
YX ₂ EGTX ₆ TSDYSX ₁₂ X ₁₃ LEX ₁₆ QAAX ₂₀ X ₂₁ FX ₂₃ X ₂₄ WLLX ₂₈ GGPSSGAPPPS (SEQ ID NO: 37)
From here;
X ₂ is Aib or A,
X ₆ is F or V,
X ₁₂ is I or Y,
X ₁₃ is Y, A, L or I;
X ₁₆ is K or E,
X ₂₀ is Q, R, E, H,
X ₂₁ is A or E,
X ₂₃ is I or V,
X ₂₄ is E, Q or N,
X ₂₈ is A or R;
compound. 4. The compound of any one of claims 1 to 3, wherein X ₁₃ LEX ₁₆ QAAX ₂₀ X ₂₁ FX ₂₃ X ₂₄ is selected from the group consisting of LLEKQAAREFIN, LLEKQAAREFIE, LLEKQAAQEFIE and LLEEQAAREFIE. The compound of claim 1 , wherein the amino acid sequence of the peptide is any one of SEQ ID NOs: 1-27 or 33-35. 6. The compound of claim 5, wherein the peptide has an amide modification at the C-terminus. The method according to any one of claims 1 to 6, wherein the substituent is

A compound selected from the group consisting of. According to claim 1, wherein the compound,

A compound selected from the group consisting of. According to claim 1, wherein the compound,

A compound selected from the group consisting of. According to claim 1, wherein the compound,

Phosphorus, compound. 11. A compound according to any one of claims 1 to 10, for use as a medicament. 12. A pharmaceutical composition comprising a compound according to any one of claims 1 to 11 for use in the prevention and/or treatment of diabetes and/or obesity. 13. Any of claims 1 to 12 for use in the prophylaxis and/or treatment of liver disorders such as hepatic steatosis, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), liver inflammation and/or fatty liver. The pharmaceutical composition according to any one of the preceding claims. A peptide having the amino acid sequence
YX ₂ EGTX ₆ TSDYSX ₁₂ X ₁₃ LEX ₁₆ QAAX ₂₀ X ₂₁ FX ₂₃ X ₂₄ WLLX ₂₈ GX ₃₀ X ₃₁ X ₃₂ X ₃₃ X ₃₄ X ₃₅ X ₃₆ X ₃₇ X ₃₈ X ₃₉ (SEQ ID NO: 47)
It has an optional amide modification at the C-terminus,
From here;
X ₂ is Aib or A,
X ₆ is F or V,
X ₁₂ is I or Y,
X ₁₃ is Y, A, L or I;
X ₁₆ is K or E,
X ₂₀ is Q, R, E, H,
X ₂₁ is A or E,
X ₂₃ is I or V,
X ₂₄ is E, Q or N,
X ₂₈ is A or R,
X ₃₀ is G or absent;
X ₃₁ is P or absent,
X ₃₂ is E, S or absent;
X ₃₃ is S, K or absent;
X ₃₄ is G or absent;
X ₃₅ is A or absent,
X ₃₆ is P or absent;
X ₃₇ is P or absent;
X ₃₈ is P or absent;
X ₃₉ is S or absent;
peptide. 15. The peptide of claim 14, wherein the amino acid sequence of the peptide is any one of SEQ ID NOs: 1-27 or 33-35.